Real-Time Biologically Inspired Action Recognition from Key Poses Using a Neuromorphic Architecture.
Intelligent agents, such as robots, have to serve a multitude of autonomous functions. Examples are, e.g., collision avoidance, navigation and route planning, active sensing of its environment, or the interaction and non-verbal communication with people in the extended reach space. Here, we focus on the recognition of the action of a human agent based on a biologically inspired visual architecture of analyzing articulated movements. The proposed processing architecture builds upon coarsely segregated streams of sensory processing along different pathways which separately process form and motion information (Layher et al., 2014). Action recognition is performed in an event-based scheme by identifying representations of characteristic pose configurations (key poses) in an image sequence. In line with perceptual studies, key poses are selected unsupervised utilizing a feature-driven criterion which combines extrema in the motion energy with the horizontal and the vertical extendedness of a body shape. Per class representations of key pose frames are learned using a deep convolutional neural network consisting of 15 convolutional layers. The network is trained using the energy-efficient deep neuromorphic networks (Eedn) framework (Esser et al., 2016), which realizes the mapping of the trained synaptic weights onto the IBM Neurosynaptic System platform (Merolla et al., 2014). After the mapping, the trained network achieves real-time capabilities for processing input streams and classify input images at about 1,000 frames per second while the computational stages only consume about 70 mW of energy (without spike transduction). Particularly regarding mobile robotic systems, a low energy profile might be crucial in a variety of application scenarios. Cross-validation results are reported for two different datasets and compared to state-of-the-art action recognition approaches. The results demonstrate, that (I) the presented approach is on par with other key pose based methods described in the literature, which select key pose frames by optimizing classification accuracy, (II) compared to the training on the full set of frames, representations trained on key pose frames result in a higher confidence in class assignments, and (III) key pose representations show promising generalization capabilities in a cross-dataset evaluation.


Serum from patients with invasive fungal infections inhibits the adherence of polymorphonuclear leukocytes and alveolar macrophages.
Adherence of various combinations of polymorphonuclear leukocytes (PMN) or alveolar macrophages (AM) and serum from nonimmunosuppressed patients wit culture-proved, invasive fungal infections or control subjects was evaluated in vitro using the standard nylon fiber pipette technique. In autologous serum, adherence of PMN from patients with wide variety of untreated fungal infections, including blastomycosis, histoplasmosis, cryptococcosis, coccidiodomycosis, and spirotrichosis, was significantly (p less than 0.001) decreased when compared with PMN from uninfected control subjects or patients with untreated bacterial infections. Studies using various combinations of PMN and serums from patients with blastomycosis suggested that the adherence defect was due to a serum disorder rather than an intrinsic cellular abnormality. Preincubation in serum from patients with blastomycosis decreased the adherence of control PMN. Preincubation in control serum corrected the decreased adherence of PMN from patients with blastomycosis. Additional studies revealed that the inhibitor was heat-stable and reversible, being present before, but not after, treatment or spontaneous resolution of the patient's infections. Adherence of AM from patients with fungal infection was also normal except when AM were preincubated in serum from patients with untreated fungal infections. We concluded that the intrinsic adherence of PMN and AM from patients with untreated fungal infections is normal, but that these patients do have an extrinsic heat-labile serum factor that can decrease the adherence of PMN and AM.


MicroRNAs and liver disease: viral hepatitis, liver fibrosis and hepatocellular carcinoma.
MicroRNAs (miRNAs), a type of highly conserved small non-coding RNAs with an average length of 22 nucleotides, may serve as major regulators of gene expression and indispensable components of cellular gene expression networks. They have critical roles in normal biological processes and have been linked to many tumours, including hepatocellular carcinoma (HCC). HCC is the fifth most common malignancy and the third leading cause of cancer-related death worldwide. Viral hepatitis, caused by infection with hepatitis B or C viruses (HBV or HCV), can increase the risk of HCC and contributes to a significant disease burden around the world. Because of the variety of molecular alterations that may arise during the development and progression of HCC, standard of care and treatment for patients with HCC remains unsatisfactory. MiRNAs have been shown to participate in the pathogenesis of both HBV and HCV. In addition to a role in pathogenesis, miRNAs have significant clinical value in the early diagnosis of HCC since they are present in the blood and can be used as diagnostic markers and potential targets for specific systemic treatment.


A gold glyco-nanoparticle carrying a Listeriolysin O peptide and formulated with Advax™ delta inulin adjuvant induces robust T-cell protection against listeria infection.
In the search for an effective vaccine against the human pathogen, Listeria monocytogenes (Listeria), gold glyconanoparticles (GNP) loaded with a listeriolysin O peptide LLO91-99 (GNP-LLO) were used to immunise mice, initially using a dendritic cell (DC) vaccine approach, but subsequently using a standard parenteral immunisation approach. To enhance vaccine immunogenicity a novel polysaccharide adjuvant based on delta inulin (Advax™) was also co-formulated with the GNP vaccine. Confirming previous results, DC loaded in vitro with GNP-LLO provided better protection against listeriosis than DC loaded in vitro using free LLO peptide. The immunogenicity of GNP-LLO loaded DC vaccines was further increased by addition of Advax™ adjuvant. However, as DC vaccines are expensive and impracticable for prophylactic use, we next asked whether the same GNP-LLO antigen could be used to directly target DC in vivo. Immunisation of mice with GNP-LLO plus Advax™ adjuvant induced LLO-specific T-cell immunity and protection against Listeria challenge. Protection correlated with an increased frequency of splenic CD4(+) and CD8(+) T cells, NK cells and CD8α(+) DC, and Th1 cytokine production (IL-12, IFN-γ, TNF-α, and MCP-1), post-challenge. Enhanced T-cell epitope recruitment post-challenge was seen in the groups that received Advax™ adjuvant. Immunisation with GNP-LLO91-99 plus Advax™ adjuvant provided equally robust Listeria protection as the best DC vaccine strategy but without the complexity and cost, making this a highly promising strategy for development of a prophylactic vaccine against listeriosis.


The effect of thermosensitive liposomal formulations on loading and release of high molecular weight biomolecules.
Thermosensitive liposomes are clinically-relevant nanocarriers which have been used to deliver chemotherapeutic agents to tumors in combination with local hyperthermia. However, the encapsulation and release of macromolecular therapeutic agents (proteins, nucleic acids, bioactive polymers) is often hindered by their instability during the liposome formation as well as by the low encapsulation efficiency. The objective of this study was to investigate the influence of the thermosensitive liposomal formulation on the encapsulation and release of low and high molecular weight hydrophilic drugs, in order to identify the key parameters to control during nanocarrier design, depending on the specific drug delivery application. Thermosensitive liposomes with different formulations were prepared through the combinations of different lipids, including dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), cholesterol (Chol), 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (P-Lyso-PC), and the PEGylated lipid distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy(PEG)-2000 (DSPE-PEG2000). The thin film hydration method was used for liposome preparation and loading of different water soluble molecules. The encapsulation efficiency and release profiles were investigated for a low molecular weight compound such as carboxyfluorescein (CF), proteins (albumin), and hydrophilic polymers which do not interact with the lipid bilayer, such as a linear dextran and a poly(ethylene glycol)-based star polymer. An optimised liposomal formulation [DPPC/P-lyso-PC/DSPE-PEG2000 90/10/4 (mol/mol) (LTSL)] was chosen for further application in encapsulating therapeutic proteins, such as lysozyme and the brain-derived neurotrophic factor (BDNF), which are recognized as drug carriers and potential therapeutic agents for kidney diseases and neurological disorders.


Ideal ward round making in neurosurgical practice.
The success of a perfect ward round lies in the role of the consultant leading the 'round making group' (RMG) as well as the hallmark of effective questioning and participation of each member. Twelve senior consultants with more than 10 years' experience in neurosurgical practice at three different university hospitals were observed during round making by a participant observer. Observations were made on the group climate of the RMG, the leadership pattern and language expressed by the clinician conducting the round and the effectiveness in his performance as a leader during clinical discussions. The group climate showed evidence of good productivity and flexibility with 92% and 75% consultants, pleasantness of climate was above average with only 50% (6/12) and poor objectivity with 42% (5/12) consultants. Forty two percent of the consultants were not always very well comprehensible, while only 50% (6/12) spoke exactly fitting the occasion. Only 33% (4/12) of the consultants used humour effectively, while 42% (5/12) spoke unnecessarily in between discussion and were poor in introducing the problems of patient to the round making group. Ward round making in neurosurgical practice needs a holistic approach with motivation, planning, leadership skills and structured curriculum to fulfill its objectives.


The role of free vitamin D and vitamin D binding protein in SARS-Cov-2 infection in children.
Many studies have discussed the effects of serum vitamin D deficiency in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) patients. This study aimed to investigate the relationship between SARS-CoV-2 infection severity and free vitamin D (FVD) and bioavailable vitamin D (BAVD) levels in children. A prospective case-control study design was used. Participants were divided into three groups based on the World Health Organization COVID-19 Clinical Progression Scale. Serum 25-hydroxyvitamin D (ng/mL), albumin (g/L), and vitamin D binding protein (ng/mL) levels were evaluated to investigate the relationship between disease severity and FVD and BAVD levels. In total, 82 participants were included in the study. Of those, 24.4% were uninfected (n = 20), 50% had a mild case of SARS-CoV-2 (n = 41), and 25.6% had a moderate case (n = 21). There was a statistically significant difference in FVD and BAVD levels between the groups (p = 0.026). Median FVD (p = 0.007, Cohen's d = 0.84) and BAVD (p = 0.007, Cohen's d = 0.86) levels were significantly higher in the mild group compared to the moderate group. FVD and BAVD metabolites were moderately positively correlated with lymphocyte counts (FVD: r = 0.437, p < 0.001; BAVD: r = 0.439, p < 0.001). This is the first study to demonstrate a relationship between SARS-CoV-2 symptom severity and FVD and BAVD levels. The relationship between FVD and BAVD levels and lymphocyte counts could play an important role in symptom severity and should be evaluated in further studies.


Hypoxic burden to guide CPAP treatment allocation in patients with obstructive sleep apnoea: a post hoc study of the ISAACC trial.
Hypoxic burden (HB) has emerged as a strong predictor of cardiovascular risk in obstructive sleep apnoea (OSA). We aimed to assess the potential of HB to predict the cardiovascular benefit of treating OSA with continuous positive airway pressure (CPAP). This was a post hoc analysis of the ISAACC trial (ClinicalTrials.gov: NCT01335087) including non-sleepy patients with acute coronary syndrome (ACS) diagnosed with OSA (apnoea-hypopnoea index ≥15 events·h-1) by respiratory polygraphy. Patients were randomised to CPAP or usual care and followed for a minimum of 1 year. HB was calculated as the total area under all automatically identified desaturations divided by total sleep time. Patients were categorised as having high or low baseline HB according to the median value (73.1%min·h-1). Multivariable Cox regression models were used to assess whether the effect of CPAP on the incidence of cardiovascular outcomes was dependent on the baseline HB level. The population (362 patients assigned to CPAP and 365 patients assigned to usual care) was middle-aged (mean age 59.7 years), overweight/obese and mostly male (84.5%). A significant interaction was found between the treatment arm and the HB categories. In the high HB group, CPAP treatment was associated with a significant reduction in the incidence of cardiovascular events (HR 0.57, 95% CI 0.34-0.96). In the low HB group, CPAP-treated patients exhibited a trend toward a higher risk of cardiovascular outcomes than those receiving usual care (HR 1.33, 95% CI 0.79-2.25). The differential effect of the treatment depending on the baseline HB level followed a dose-response relationship. In non-sleepy ACS patients with OSA, high HB levels were associated with a long-term protective effect of CPAP on cardiovascular prognosis.


Estimation of genetic variation in residual variance in female and male broiler chickens.
In breeding programs, robustness of animals and uniformity of end product can be improved by exploiting genetic variation in residual variance. Residual variance can be defined as environmental variance after accounting for all identifiable effects. The aims of this study were to estimate genetic variance in residual variance of body weight, and to estimate genetic correlations between body weight itself and its residual variance and between female and male residual variance for broilers. The data sets comprised 26 972 female and 24 407 male body weight records. Variance components were estimated with ASREML. Estimates of the heritability of residual variance were in the range 0.029 (s.e. = 0.003) to 0.047 (s.e. = 0.004). The genetic coefficients of variation were high, between 0.35 and 0.57. Heritabilities were higher in females than in males. Accounting for heterogeneous residual variance increased the heritabilities for body weight as well. Genetic correlations between body weight and its residual variance were -0.41 (s.e. = 0.032) and -0.45 (s.e. = 0.040), respectively, in females and males. The genetic correlation between female and male residual variance was 0.11 (s.e. = 0.089), indicating that female and male residual variance are different traits. Results indicate good opportunities to simultaneously increase the mean and improve uniformity of body weight of broilers by selection.


Effective radiovirotherapy for malignant gliomas by using oncolytic measles virus strains encoding the sodium iodide symporter (MV-NIS).
Engineered measles virus (MV) strains deriving from the vaccine lineage represent a promising oncolytic platform and are currently being tested in phase I trials. In this study, we have demonstrated that MV strains genetically engineered to express the human sodium iodide symporter (NIS) have significant antitumor activity against glioma lines and orthotopic xenografts; this compares favorably with the MV strain expressing the human carcinoembryonic antigen, which is currently in clinical testing. Expression of NIS protein in infected cells results in effective concentration of radioactive iodine, which allows for in vivo monitoring of localization of MV-NIS infection by measuring uptake of (123)I or (99m)Tc. In addition, radiovirotherapy with MV-NIS followed by (131)I administration resulted in significant increase of MV-NIS antitumor activity as compared with virus alone in both subcutaneous (p=0.0003) and orthotopic (p=0.004) glioblastoma models. In conclusion, MV-NIS-based radiovirotherapy has significant antitumor activity against glioblastoma multiforme and represents a promising candidate for clinical translation.


Annexins as disease modifiers.
The annexins are a family of calcium-dependent phospholipid binding proteins which are present in all eukaryotes. There are currently 12 identified human annexins all of which contain unique calcium binding sites, encoded in the highly conserved annexin repeat motifs within the C terminal core. In addition to the C terminal core the annexins contain a significantly more variable N terminal head. It is this domain which endows each annexin with unique functions in a diverse range of cellular processes including; endo- and exocytosis, cytoskeletal regulation and membrane conductance and organisation. Given their involvement in such a variety of processes it is not surprising that the annexins have also been implicated in a range of disease pathologies. Although there is no singular disease state directly attributed to a dysregulation in annexin function, several pathological conditions are suggested to be modified by the annexins. In this review we shall focus on the growing evidence for the role of the annexins in the progression of cancer, diabetes and the autoimmune disorder anti-phospholipid syndrome.


Cry toxin specificities of insect ABCC transporters closely related to lepidopteran ABCC2 transporters.
In this study, we examined insect and human ABCC transporters closely related to the lepidopteran ABC transporter C2 (ABCC2), a powerful receptor for the Bacillus thuringiensis Cry toxin, for their responses to various Cry toxins. ABCC2 and the lepidopteran ABC transporter C3 (ABCC3) conferred cultured cells with susceptibility to a lepidopteran-specific Cry1Aa toxin but not to lepidopteran-specific Cry1Ca and Cry1Da. One coleopteran ABCC transporter specifically responded to a coleopteran-specific Cry8Ca toxin. ABCC transporters from a dipteran insect and humans did not respond to any of the tested Cry toxins that are active to lepidopteran and coleopteran insects. These results yield important information for our understanding of insect specificity of Cry toxins and provide the first demonstration of a coleopteran ABCC transporter that serves as a Cry toxin receptor.


Cost-effective shock wave hydrophones.
This article presents the design of a newly developed wideband, quantitative shock wave sensor (hydrophone) made of piezoelectric material polyvinylidene difluoride (PVDF). The novel aspects of this device include a disposable sensor element to reduce the per shot costs, and a self-monitoring feature that informs the user that the sensor needs to be replaced. The development, design, and use of the sensors are presented. Results indicate that these sensors can fulfill the requirements for accurate, wideband measurement of lithotripter fields, suitable for quantitative biophysical research or Food and Drug Administration submissions. The disposable feature of the hydrophone system makes the system cost effective for research, quality assurance, and regulatory submissions.


Converging on the function of intrinsically disordered nucleoporins in the nuclear pore complex.
Several biological mechanisms involve proteins or proteinaceous components that are intrinsically disordered. A case in point pertains to the nuclear pore complex (NPC), which regulates molecular transport between the nucleus and the cytoplasm. NPC functionality is dependent on unfolded domains rich in Phe-Gly (FG) repeats (i.e., FG-domains) that collectively act to promote or hinder cargo translocation. To a large extent, our understanding of FG-domain behavior is limited to in vitro investigations given the difficulty to resolve them directly in the NPC. Nevertheless, recent findings indicate a collective convergence towards rationalizing FG-domain function. This review aims to glean further insight into this fascinating problem by taking an objective look at the boundary conditions and contextual details underpinning FG-domain behavior in the NPC. Here, we treat the FG-domains as being commensurate with polymeric chains to address ambiguities such as for instance, how FG-domains tethered to the central channel of the NPC would behave differently as compared with their free-floating counterparts in solution. By bringing such fundamental questions to the fore, this review seeks to illuminate the importance of how such parameters can hold influence over the structure-function relation of intrinsically disordered proteins in the NPC and beyond.


Automatic normative quantification of brain tissue volume to support the diagnosis of dementia: A clinical evaluation of diagnostic accuracy.
To assesses whether automated brain image analysis with quantification of structural brain changes improves diagnostic accuracy in a memory clinic setting. In 42 memory clinic patients, we evaluated whether automated quantification of brain tissue volumes, hippocampal volume and white matter lesion volume improves diagnostic accuracy for Alzheimer's disease (AD) and frontotemporal dementia (FTD), compared to visual interpretation. Reference data were derived from a dementia-free aging population (n = 4915, aged >45 years), and were expressed as age- and sex-specific percentiles. Experienced radiologists determined the most likely imaging-based diagnosis based on structural brain MRI using three strategies (visual assessment of MRI only, quantitative normative information only, or a combination of both). Diagnostic accuracy of each strategy was calculated with the clinical diagnosis as the reference standard. Providing radiologists with only quantitative data decreased diagnostic accuracy both for AD and FTD compared to conventional visual rating. The combination of quantitative with visual information, however, led to better diagnostic accuracy compared to only visual ratings for AD. This was not the case for FTD. Quantitative assessment of structural brain MRI combined with a reference standard in addition to standard visual assessment may improve diagnostic accuracy in a memory clinic setting.


Selfing rates vary with floral display, pollinator visitation and plant density in natural populations of Mimulus ringens.
Variation in selfing rates within and among populations of hermaphroditic flowering plants can strongly influence the evolution of reproductive strategies and the genetic structure of populations. This intraspecific variation in mating patterns may reflect both genetic and ecological factors, but the relative importance of these factors remains poorly understood. Here, we explore how selfing in 13 natural populations of the perennial wildflower Mimulus ringens is influenced by (a) pollinator visitation, an ecological factor, and (b) floral display, a trait with a genetic component that also responds to environmental variation. We also explore whether genetically based floral traits, including herkogamy, affect selfing. We found substantial variation among populations in selfing rate (0.13-0.55). Selfing increased strongly and significantly with floral display, among as well as within populations. Selfing also increased at sites with lower pollinator visitation and low plant density. However, selfing was not correlated with floral morphology. Overall, these results suggest that pollinator visitation and floral display, two factors that interact to affect geitonogamous pollinator movements, can influence the selfing rate. This study identifies mechanisms that may play a role in maintaining selfing rate variation among populations.


Effects of Participation and Collaboration on Perceived Effectiveness of Core Public Health Functions.
We measured the perceived effectiveness of core public health services at the community level, and we assessed whether certain system factors were associated with perceived effectiveness. We used data from the National Longitudinal Survey of Local Public Health Systems, conducted in 1998 and 2006, to examine the effects of the contributions of health departments and the participation of other agencies to core functions on the perceived effectiveness of community delivery of core assessment, policy development, and assurance functions. Perceived effectiveness increased over time for all 3 core function areas (range = 41%-53% in 2006). Multivariable results showed that local health department contribution to effort was positively associated with perceived effectiveness of public health core functions. The significance of participation of individual types of agencies or organizations varied by core function areas, except for local government agencies and hospitals, which were significantly positively associated with all 3 core function areas. We furthered our understanding of the significance of the contributions and participations of local health departments and of other individual agencies or organizations in the perceived effectiveness of delivery of core functions.


Oxyphylla A Promotes Degradation of α-Synuclein for Neuroprotection via Activation of Immunoproteasome.
Parkinson's disease (PD), the second most common neurodegenerative disorder, is neuropathologically characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta (SNc) and the presence of Lewy bodies in surviving neurons. α-synuclein (α-syn) is the major component of Lewy bodies and its deposition in neurons is critical pathological event in the pathogenesis of PD. Herein, we reported that Oxyphylla A, a novel lead compound from the fruit of Alpinia oxyphylla, significantly promoted α-syn degradation in a cellular PD model. When exploring the molecular pathways, we found that Oxyphylla A promoted α-syn degradation in a ubiquitin proteasome system (UPS)-dependent and autophagy-independent manner. We further confirmed that Oxyphylla A enhanced UPS activity by upregulating 20S subunit PSMB8 expression. A mechanism study revealed that Oxyphylla A activated the PKA/Akt/mTOR pathway to trigger PSMB8 expression and enhance UPS activity. Finally, we illustrated that Oxyphylla A alleviated the accumulation of both Triton-soluble and Triton-insoluble forms of α-syn and protected against α-syn-induced neurotoxicity in A53T α-syn transgenic mice. These findings suggest that the activation of UPS, via small molecular UPS enhancers including Oxyphylla A, may be a therapeutic strategy for intervention against PD and related diseases.


Differential contributions of ovarian and extraovarian factors to age-related reductions in plasma estradiol and progesterone during the estrous cycle of C57BL/6J mice.
The relative contributions of ovarian and extra-ovarian factors to the altered ovarian steroidal profiles of middle-aged mice were assessed by reciprocal, heterochronic ovarian grafting. Ovaries from cycling, young (2 months), and middle-aged (12 months) mice were exchanged by grafting under the renal capsules. Blood samples were obtained daily at midday throughout the estrous cycle for measurement of estradiol (E2) and 3-4 h after lights-out on proestrus to measure the preovulatory elevation of progesterone (P4). Middle-aged intact mice had lower mean concentrations of E2 during the cycle, no detectable midday preovulatory elevation of E2, and an attenuated preovulatory increase of P4 compared to young mice. Ovarian grafts from young donors failed to increase mean E2 levels of middle-aged mice, but did restore the preovulatory elevation of E2 and preovulatory P4 to levels of young controls. Reciprocal grafting confirmed these findings: ovaries from middle-aged donors in young hosts produced mean E2 levels equivalent to those of young mice but were unable to support a preovulatory increase of E2 or a preovulatory P4 level equivalent to that of young controls. These results reveal differential contributions of ovarian and extra-ovarian factors to age changes in E2 and P4. They indicate that ovarian aging plays an important role in attenuating the preovulatory increase of E2 and P4, but extra-ovarian, presumably neuroendocrine, age changes underlie the mean reduction of E2 levels across the estrous cycle.


[Main trends in the demographic development in Russia and dynamics of the rate of patients with schizophrenia].
The authors analyze both the demographic situation in Russia during the past years and its perspective. A number of patients with paranoid schizophrenia and with schizophrenia on the whole (all forms of disease) was estimated for the period of the coming 15 years (till 2011 yr.). The level of the patients' detection in some Russian regions was estimated by means of comparison of the calculated number of the patients (according to tables of incidence) and actually known during the same period. The proportion of registered schizophrenic patients was up to 95.7% from calculated data in Moscow, while in other regions this index was not more than 50%. A low detection of the schizophrenic patients was explained by both specificity of the organization of psychiatric care in the regions, a shortage of psychiatrists as well as by other factors.


Assessment of fluorescence in situ hybridization and hybrid capture 2 analyses of cervical cytology specimens diagnosed as low grade squamous intraepithelial lesion for the detection of high grade cervical intraepithelial neoplasia.
To assess Hybrid Capture 2 (HC2) and fluorescence in situ hybridization (FISH) for the detection of cervical intraepithelial neoplasia 2 or worse (CIN 2+) in patients with a cytologic diagnosis of low grade squamous intraepithelial lesion (LSIL). Residual samples from 115 LSIL-diagnosed cervical cytology specimens were evaluated by high-risk human papillomavirus (HR-HPV) HC2 testing and FISH using biotin-labeled probes to HR-HPV and chromosomal probes to 3q26 (TERC) and 8q24 (CMYC). A cervical biopsy diagnosis of CIN 2+ was considered as evidence of high grade disease. The positive and negative predictive values of HC2 and FISH for detecting patients with CIN 2+ were 32% vs. 37% and 100% vs. 93%, respectively. The sensitivities of HC2 and FISH for CIN 2+ were not significantly different (100% vs. 90%, p = 0.25), while the specificity of HC2 was significantly lower than that of FISH (28% vs. 48%, p=0.003). FISH diagnosed fewer specimens as positive as compared to HC2 (62% vs. 79%). These preliminary data suggest that FISH testing may be useful for determining which patients with LSIL are most likely to have CIN 2+ on clinical follow-up.


Synthesis and helical structures of poly(ω-alkynamide)s having chiral side chains: effect of solvent on their screw-sense inversion.
New ω-alkynamides, (S)-HC≡CCH2CONHCH2CH(CH3)CH2CH3 (1) and (S)-HC≡CCH2CH2CONHCH(CH3)CH2CH2CH2CH2CH3 (2) were synthesized and polymerized with a rhodium catalyst in CHCl3 to obtain cis-stereoregular poly(ω-alkynamide)s (poly(1) and poly(2)). Polarimetric, CD, and IR spectroscopic studies revealed that in solution the polymers adopted predominantly one-handed helical structures stabilized by intramolecular hydrogen bonds between the pendent amide groups. This behavior was similar to that of the corresponding poly(N-alkynylamide) counterparts (poly(3) and poly(4)) reported previously, whereas the helical senses were opposite to each other. The helical structures of the poly(ω-alkynamide)s were stable upon heating similar to those of the poly(N-alkynylamide)s, but the solvent response was completely different. An increase in MeOH content in CHCl3/MeOH resulted in inversion of the predominant screw-sense for poly(1) and poly(2). Conversely, poly(3) was transformed into a random coil, and poly(4) maintained the predominant screw-sense irrespective of MeOH content. The solvent dependence of predominant screw-sense for poly(1) and poly(2) was reasonably explained by molecular orbital studies using the conductor-like screening model (COSMO).


Sequence-specific interaction of Hoechst 33258 with the minor groove of an adenine-tract DNA duplex studied in solution by 1H NMR spectroscopy.
The interaction of Hoechst 33258 with the minor groove of the adenine-tract DNA duplex d(CTTTTGCAAAAG)2 has been studied in both D2O and H2O solutions by 1D and 2D 1H NMR spectroscopy. Thirty-one nuclear Overhauser effects between drug and nucleotide protons within the minor groove of the duplex, together with ring-current induced perturbations to the chemical shifts of basepair and deoxyribose protons, define the position and orientation of the bound dye molecules. Two drug molecules bind cooperatively and in symmetry related orientations at the centre of the 5'-TTTT and 5'-AAAA sequences with the binding interactions spanning only the four A-T basepairs. The positively charged N-methylpiperazine moieties point towards the centre of the duplex while the phenol groups are disposed towards the 3'-ends of the sequence. Resonance averaging is apparent for both the D2/D6 and D3/D5 phenol protons and D2"'/D6"' and D3"'/D5"' of the N-methylpiperazine ring and is consistent with these groups being involved in rapid rotation or ring-flipping motions in the bound state. Interstrand NOEs between adenine H2s and deoxyribose H1' are consistent with a high degree of propeller twisting of the A-T basepairs at the binding site of the aromatic benzimidazole and phenol rings of Hoechst. The data imply that the minor groove is particularly narrow with many contacts between the complementary curved surfaces of the drug and DNA indicating that strong van der Waals interactions, involving the floor and the walls of the minor groove, stabilize the complex. In our model the NH groups of the benzimidazole rings are positioned to make a pair of bifurcated hydrogen bonds with the adenine N3 and thymine O2 on the floor of the minor groove.


Evaluating the sources and functions of gradiency in phoneme categorization: An individual differences approach.
During spoken language comprehension listeners transform continuous acoustic cues into categories (e.g., /b/ and /p/). While long-standing research suggests that phonetic categories are activated in a gradient way, there are also clear individual differences in that more gradient categorization has been linked to various communication impairments such as dyslexia and specific language impairments (Joanisse, Manis, Keating, & Seidenberg, 2000; López-Zamora, Luque, Álvarez, & Cobos, 2012; Serniclaes, Van Heghe, Mousty, Carré, & Sprenger-Charolles, 2004; Werker & Tees, 1987). Crucially, most studies have used 2-alternative forced choice (2AFC) tasks to measure the sharpness of between-category boundaries. Here we propose an alternative paradigm that allows us to measure categorization gradiency in a more direct way. Furthermore, we follow an individual differences approach to (a) link this measure of gradiency to multiple cue integration, (b) explore its relationship to a set of other cognitive processes, and (c) evaluate its role in individuals' ability to perceive speech in noise. Our results provide validation for this new method of assessing phoneme categorization gradiency and offer preliminary insights into how different aspects of speech perception may be linked to each other and to more general cognitive processes. (PsycINFO Database Record


Isolation and characterization of the nuclear serpin MENT.
A balance between proteolytic activity and protease inhibition is required to maintain the appropriate function of biological systems in which proteases play a role. The Myeloid and Erythroid Nuclear Termination protein, MENT, is a nonhistone heterochromatin-associated serpin that is an effective inhibitor of the papain-like cysteine proteases. Our laboratories have extensively investigated the dual functions of this protein, namely, chromatin condensation and protease inhibition. Unlike other serpins to date, MENT contains a unique insertion between the C- and D-helices known as the "M-loop." This loop contains two critical functional motifs that allow the nuclear function of MENT, namely, nuclear localization and DNA binding. However, the nuclear function of MENT is not restricted to the activities of the M-loop alone. In vitro, MENT brings about the dramatic remodeling of chromatin into higher-order structures by forming protein bridges via its reactive center loop. Further, we have determined that in a protease-mediated effect, DNA can act as a cofactor to accelerate the rate at which MENT can inhibit its target proteases. In this chapter, we discuss the isolation of MENT from native chicken blood as well as recombinant protein produced in Escherichia coli. Various techniques including in vitro functional assays and biophysical characterization are explained that can be used to elucidate the ability of the protein to interact with DNA and other deoxynucleoprotein complexes. In situ chromatin precipitation using natively purified MENT is also detailed.


[Central case management in surgery].
Up to now, in most surgical departments there has been no central patient management coordinating the admission, the diagnostic work-up, and the surgical procedures of patients with the resources available in the hospital. In future, however, it will be essential for surgical departments to establish such a central patient management, not only in view of the importance of patient-oriented medicine but also considering the planned introduction of diagnosis related groups. A central patient management will reduce preoperative and overall length of stay through adequate organisation and communication structures. In addition, a central patient management will make optimal use of the available resources in terms of operation rooms and hospital beds. In this article, we will present our basic concept of a central patient management in a surgical university department.


Leukoplakia and angiopoiesis resistance of Herba Erigerontis.
To evaluate the effects of Herba Erigerontis on angiopoiesis and carcinogenesis; to seek an effective drug for prevention of leukoplakia progression to carcinoma and to stop the mechanism of angiopoiesis. Carcinomatous conversions of golden hamster cheek pouches induced by Salley way and inhibited by Herba Erigerontis. Ink perfusion, analysis of images, resin cast form of micro-vessel, a-SMA detection and histopathological examination were applied to observe their changing patterns. The rate of leukoplakia conversion to tumor of the Herba Erigerontis group was half that of model group. The proportion of normal cells was approximately 4 times greater than that of model. The a-SMA levels of the Herba Erigerontis treated group was higher than that of model (P < 0.001). There was no significant difference between angeion vessel area and density (P > 0.05) in these 2 groups, which were elevated compared to the control. Nevertheless, the resin casts of the microvasculature in the Herba Erigerontis group showed that its configuration and spatial arrangement was similar to the control group. Herba Erigerontis has the function of inhibiting leukoplakia progression to tumor. This may be due to its ability to upregulate the expression of a-SMA. It has no obvious effect on angio-hyperplasia and expansion in the course of leukoplakia progression to tumor. However, it may preserve the angio-configuration and spatial arrangement and keep intact the angeion vessel wall. Its mechanism of promoting blood circulation may be through removing blood stasis and benign angiopoiesis so as to resist carcinogenesis.


Heteromeric three-stranded coiled coils designed using a Pb(II)(Cys)3 template mediated strategy.
Three-stranded coiled coils are peptide structures constructed from amphipathic heptad repeats. Here we show that it is possible to form pure heterotrimeric three-stranded coiled coils by combining three distinct characteristics: (1) a cysteine sulfur layer for metal coordination, (2) a thiophilic, trigonal pyramidal metalloid (Pb(II)) that binds to these sulfurs and (3) an adjacent layer of reduced steric bulk generating a cavity where water can hydrogen bond to the cysteine sulfur atoms. Cysteine substitution in an a site yields Pb(II)A2B heterotrimers, while d sites provide pure Pb(II)C2D or Pb(II)CD2 scaffolds. Altering the metal from Pb(II) to Hg(II) or shifting the relative position of the sterically less demanding layer removes heterotrimer specificity. Because only two of the eight or ten hydrophobic layers are perturbed, catalytic sites can be introduced at other regions of the scaffold. A Zn(II)(histidine)3(H2O) centre can be incorporated at a remote location without perturbing the heterotrimer selectivity, suggesting a unique strategy to prepare dissymmetric catalytic sites within self-assembling de novo-designed proteins.


Small noncoding RNAs encoded within the bovine herpesvirus 1 latency-related gene can reduce steady-state levels of infected cell protein 0 (bICP0).
Following acute infection in mucosal epithelium, bovine herpes virus 1 (BHV-1) establishes lifelong latency in sensory neurons within trigeminal ganglia. The latency-related RNA (LR-RNA) is abundantly expressed in sensory neurons of latently infected calves. Expression of LR proteins is necessary for the latency reactivation cycle because a mutant virus that does not express LR proteins is unable to reactivate from latency after dexamethasone treatment. LR-RNA sequences also inhibit bICP0 expression, productive infection, and cell growth. However, it is unclear how LR-RNA mediates these functions. In this study, we identified a 463-bp region within the LR gene (the XbaI-PstI [XP] fragment) that inhibited bICP0 protein and RNA expression in transiently transfected mouse neuroblastoma cells. Small noncoding RNAs (sncRNAs) encoded within the XP fragment (20 to 90 nucleotides in length) were detected in transiently transfected mouse neuroblastoma cells. Two families of sncRNAs were cloned from this region, and each family was predicted to contain a mature microRNA (miRNA). Both miRNAs were predicted to base pair with bICP0 mRNA sequences, suggesting that they reduce bICP0 levels. To test this prediction, sequences encompassing the respective sncRNAs and mature miRNAs were synthesized and cloned into a small interfering RNA expression vector. Both sncRNA families and their respective miRNAs inhibited bICP0 protein expression in mouse neuroblastoma cells and productive infection in bovine cells. In trigeminal ganglia of latently infected calves, an sncRNA that migrated between nucleotides 20 and 25 hybridized to the XP fragment. During dexamethasone-induced reactivation from latency, XP-specific sncRNA levels were reduced, suggesting that these sncRNAs support the establishment and maintenance of lifelong latency in cattle.


Spontaneous autoresuscitation in a model of respiratory control.
We introduce a closed-loop model of respiratory control incorporating a conductance-based central pattern generator (CPG), low-pass filtering of CPG output by the respiratory musculature, gas exchange in the lung, metabolic oxygen demand, and chemosensation. The CPG incorporates Butera, Rinzel and Smith (BRS)'s (1999) conditional pacemaker model. BRS model cells can support quiescent, bursting, or beating activity depending on the level of excitatory drive; we identify these activity modes with apnea (cessation of breathing), eupnea (normal breathing), and tachypnea (excessively rapid breathing). We demonstrate the coexistence of two dynamically stable behaviors in the closed-loop model, corresponding respectively to eupnea and tachypnea. The latter state represents a novel failure mode within a respiratory control model. In addition, the closed-loop system exhibits a form of autoresuscitation: conductances intrinsic to the BRS model buffer the CPG against brief episodes of hypoxia, steering the system away from catastrophic collapse as can occur with tachypnea.


Cutaneous Alternaria infectoria infection in a dog in association with therapeutic immunosuppression for the management of immune-mediated haemolytic anaemia.
A 4-year-old, ovariohysterectomized, English springer spaniel on immunosuppressive therapy was re-examined for the review of its immune-mediated haemolytic anaemia and the recent development of skin lesions. For the 3 months since hospital discharge, the dog had been receiving 1.3 mg/kg prednisolone and 2.6 mg/kg ciclosporin, both administered orally twice daily. Physical examination revealed hepatomegaly and multiple, purulent, crusting, erosive to ulcerative lesions over different body areas. Onychorrhexis had occurred on one digit and the underlying corium had blackened. There were two proliferative and one plaque-like lesions in the mouth. Thick walled fungal hyphae were detected in impression smears from all skin lesions and staining with periodic acid-Schiff's stain confirmed the presence of multiple fungal hyphae and spores in all biopsies examined. Fungal culture isolated a heavy, pure growth of an Alternaria sp. which was identified as A. infectoria by sequencing the internal transcribed spacer 1 region of the rRNA gene. The animal's condition prevented detailed investigation of the oral lesions. Withdrawal of the ciclosporin and reduction of the prednisolone dosage resulted in spontaneous resolution of the skin lesions within 40 days. Further gradual decrements in the prednisolone dosage to zero were carried out without recurrence of the immune-mediated haemolytic anaemia. After 12 months, there has been no recurrence of either the skin lesions or the anaemia. To the authors' knowledge, this is the first reported case of A. infectoria infection in a dog.


Cognitive-behavioral therapy, behavioral weight loss, and sequential treatment for obese patients with binge-eating disorder: a randomized controlled trial.
Cognitive-behavioral therapy (CBT) is the best established treatment for binge-eating disorder (BED) but does not produce weight loss. The efficacy of behavioral weight loss (BWL) in obese patients with BED is uncertain. This study compared CBT, BWL, and a sequential approach in which CBT is delivered first, followed by BWL (CBT + BWL). 125 obese patients with BED were randomly assigned to 1 of the 3 manualized treatments delivered in groups. Independent assessments were performed posttreatment and at 6- and 12-month follow-ups. At 12-month follow-up, intent-to-treat binge-eating remission rates were 51% (CBT), 36% (BWL), and 40% (CBT + BWL), and mean percent BMI losses were -0.9, -2.1, and 1.5, respectively. Mixed-models analyses revealed that CBT produced significantly greater reductions in binge eating than BWL through 12-month follow-up and that BWL produced significantly greater percent BMI loss during treatment. The overall significant percent BMI loss in CBT + BWL was attributable to the significant effects during the BWL component. Binge-eating remission at major assessment points was associated significantly with greater percent BMI loss cross-sectionally and prospectively (i.e., at subsequent follow-ups). CBT was superior to BWL for producing reductions in binge eating through 12-month follow-up, while BWL produced statistically greater, albeit modest, weight losses during treatment. Results do not support the utility of the sequential approach of providing BWL following CBT. Remission from binge eating was associated with significantly greater percent BMI loss. Findings support BWL as an alternative treatment option to CBT for BED.


Silicon inhibits cadmium uptake by regulating the genes associated with the lignin biosynthetic pathway and plant hormone signal transduction in maize plants.
Cadmium (Cd) contamination in soil poses a severe threat to plant growth and development. In contrast, silicon (Si) has shown promise in enhancing plant resilience under Cd-induced stress. In this study, we conducted an integrated investigation employing morphological studies, gene expression analysis, and metabolomics to unravel the molecular mechanisms underlying Cd tolerance in maize plants. Our results demonstrate that Si biofortification significantly mitigated Cd stress by reducing Cd accumulation in plant tissues, increasing Si content, and enhancing maize biomass in Cd-stressed plants resulted in a substantial enhancement in shoot dry weight (+ 75%) and root dry weight (+ 30%). Notably, Si treatment upregulated key lignin-related genes (TaPAL, TaCAD, Ta4CL, and TaCOMT) and promoted the accumulation of metabolites (sinapyl alcohol, phenylalanine, p-coumaryl alcohol, cafeyl alcohol, and coniferaldehyde) essential for cell wall strength, particularly under Cd stress conditions. Si application enriched the signal transduction by hormones and increased resistance by induction of biosynthesis genes (TaBZR1, TaLOX3, and TaNCDE1) and metabolites (brassinolide, abscisic acid, and jasmonate) in the roots and leaves under Cd stress. Furthermore, our study provides a comprehensive view of the intricate molecular crosstalk between Si, Cd stress, and plant hormonal responses. We unveil a network of genetic and metabolic interactions that culminate in a multifaceted defense system, enabling maize plants to thrive even in the presence of Cd-contaminated soil. This knowledge not only advances our understanding of the protective role of Si but also highlights the broader implications for sustainable agricultural practices. By harnessing the insights gained from this research, we may pave the way for innovative strategies to fortify crops against environmental stressors, ultimately contributing to the goal of food security in an ever-changing world. In summary, our research offers valuable insights into the protective mechanisms facilitated by Si, which enhance plants' ability to withstand environmental stress, and holds promise for future applications in sustainable agriculture.


Effects of recognizing depression with a standardized questionnaire (CES-D) versus patient reporting of depression after a single-standardized question on the outcomes of treatment for hepatitis C with pegylated interferon-α-2b and ribavirin.
Depression may worsen during antiviral treatment for hepatitis C virus, resulting in noncompliance treatment. The aim was to compare the response and compliance rates between the groups of veteran patients using two different methods of identifying depression, either the Centers for Epidemiology Studies for Depression Scale (group A) questionnaire or the report of symptoms of depression after a single-standardized question by the health care provider (group B). One hundred and twenty-nine patients were randomly assigned to the two groups before the treatment. No statistical differences were noted in baseline characteristics between the groups. Depression was common in both the groups. No difference between initial Centers for Epidemiologic Studies Depression Scale scores and diagnosis of depression between the two groups was noted. Furthermore, the number of patients diagnosed with depression during the treatment was similar in each group. There were no significant differences between the groups in rates of sustained viral response (30% group A, 35% group B) or in rates of overall compliance with patients receiving more than 90% of prescribed PegIntronA therapy (44% group A, 39% group B), and ribavirin (32% group A and 37% group B). The use of the Centers for Epidemiology Studies for Depression Scale questionnaire to recognize depression had no significant advantage over patient reporting of depression symptoms after a single-standardized question on the hepatitis C virus clearance and the treatment compliance rates in veteran populations.


Substituting redberry juniper for oat hay in lamb feedlot diets: Carcass characteristics, adipose tissue fatty acid composition, and sensory panel traits.
We hypothesized that ground juniper and dried distillers grains with solubles (DDGS) fed to lambs would have no negative effects on end products. Lambs were individually fed diets containing mainly sorghum grain and oat hay (control) or DDGS-based diets consisting mainly of DDGS, sorghum grain, and oat hay (0JUN), or 33% (33JUN), 66% (66JUN), or 100% (100JUN) of the hay replaced by juniper. Lambs fed control had greater (P<0.03) shrunk BW and HCW vs. lambs fed 0JUN, but other carcass characteristics and sensory panel traits were similar (P>0.23). As juniper increased in the DDGS-based diets, HCW increased quadratically (P=0.01) and LM fatty acid composition was altered. Feeding juniper did not negatively affect (P>0.57) off-flavor in chops, but enhanced (linear, P<0.05) juiciness, tenderness, and flavor intensity. Lambs fed diets with a combination of oat hay and ground juniper appeared to produce the most acceptable carcasses and lamb chops.


Preferable Transplant Site for Hepatocyte Transplantation in a Rat Model.
Intraportal injection is regarded as the current standard procedure of hepatocyte transplantation (HTx). In islet transplantation, which shares many aspects with HTx, recent studies have clarified that instant blood-mediated inflammatory reaction (IBMIR), characterized by strong innate immune responses, can cause poor engraftment, so other transplant sites to avoid such a reaction have been established. Although IBMIR was reported to occur in HTx, few reports have evaluated alternative transplant sites for HTx. In this study, we sought to determine the optimum transplant site for HTx. Rat hepatocytes (1.0 × 107) were transplanted at the 9 transplant sites (intraportal (IPO), intrasplenic (IS), liver parenchyma, subcutaneous, intraperitoneal, renal subcapsular, muscle, inguinal subcutaneous white adipose tissue, and omentum) of analbuminemic rats. The serum albumin levels, immunohistochemical staining (albumin, TUNEL, and BrdU), and in vivo imaging of the grafts were evaluated. The serum albumin levels of the IPO group were significantly higher than those of the other groups (p < .0001). The BrdU-positive hepatocyte ratio of liver in the IS group (0.9% ± 0.2%) was comparable to that of the IPO group (0.9% ± 0.3%) and tended to be higher than that of the spleen in the IS group (0.5% ± 0.1%, p = .16). Considering the in vivo imaging evaluation and the influence of splenectomy, the graft function in the IS group may be almost entirely achieved by hepatocytes that have migrated to the liver. The present study clearly showed that the intraportal injection procedure is more efficient than other procedures for performing HTx.


Cyclic and Torsional Fatigue Resistance of Reciprocating Single Files Manufactured by Different Nickel-titanium Alloys.
The aim of this study was to evaluate the cyclic and torsional fatigue resistance of the following reciprocating single-file systems: ProDesign R 25.06 (Easy Equipamentos Odontológicos, Belo Horizonte, Brazil), Reciproc R25 (VDW GmbH, Munich, Germany), and Unicone L25 (Medin SA, Nové Město in Moravě, Czech Republic). Sixty instruments of the ProDesign R, Reciproc R25, and Unicone L25 systems (n = 20) were used. Cyclic fatigue resistance was tested measuring the time to failure in an artificial stainless steel canal with a 60° angle and a 5-mm radius of curvature (n = 10). Torque and angle of rotation at failure of new instruments (n = 10) in the 3 mm from the tip portion were measured during torsional testing according to ISO 3630-1. The fractured surface of each fragment was examined by scanning electron microscopy. Data were analyzed using 1-way analysis of variance and Tukey tests, and the level of significance was set at 5%. The cyclic fatigue resistance values of ProDesign R 25.06 were significantly higher than the other groups (P < .05). Reciproc R25 showed higher fatigue resistance than Unicone L25 (P < .05). In relation to the torsional test, the ProDesign R 25.06 and Unicone L25 systems showed higher angular rotation until fracture than Reciproc R25 (P < .05). However, Reciproc R25 and Unicone L25 showed higher torque load than ProDesign R 25.06 (P < .05). Scanning electron microscopic analysis showed similar and typical features of cyclic and torsional failure for all instruments tested. ProDesign R presented the highest cyclic fatigue resistance and angular rotation to failure compared with Reciproc and Unicone. However, Reciproc showed higher torsional strength to failure.


Function of microRNA-375 and microRNA-124a in pancreas and brain.
In recent years, our understanding of how gene regulatory networks control cell physiology has improved dramatically. Studies have demonstrated that transcription is regulated not only by protein factors, but also by small RNA molecules, microRNAs (miRNAs). The first miRNA was discovered in 1993 as a result of a genetic screen for mutations in Caenorhabditis elegans. Since then, the use of sophisticated techniques and screening tools has promoted a more definitive understanding of the role of miRNAs in mammalian development and diseases. miRNAs have emerged as important regulators of genes involved in many biological processes, including development, cell proliferation and differentiation, apoptosis and metabolism. Over the last few years, the number of reviews dealing with miRNAs has increased at an impressive pace. In this review, we present general information on miRNA biology and focus more closely on comparing the expression, regulation and molecular functions of the two miRNAs, miR-375 and miR-124a. miR-375 and miR-124a share similar features; they are both specifically expressed in the pancreas and brain and directly bind a common target gene transcript encoding myotrophin, which regulates exocytosis and hormone release. Here, we summarize the available data obtained by our group and other laboratories and provide an overview of the specific molecular function of miR-375 and miR-124a in the pancreas and the brain, revealing a potential functional overlap for these two miRNAs and the emerging therapeutic potential of miRNAs in the treatment of human metabolic diseases.


Studying Local Electrostatics by Terahertz Spectroscopy Using Amines as a Probe.
In a previous study[1] we could show that a large amplitude mode of the zwitterion glycine can serve as a sensitive probe for protonation and allows to deduce local pKa values. Here we show that the underlying concept is more general: We present the results of a pH dependent measurement of Terahertz-FTIR (THz-FTIR) spectra of solvated amines, i. e. Diethylamine (DEA), Triethylamine (TEA), and Diisopropylamine (DiPA). We show that amines serve as a sensitive, label free probe for local protonation. Protonation of the amines yield intensity changes which can be quantified by precise THz spectroscopy (30 cm-1 -450 cm-1 ). A detailed analysis allows us to correlate the titration spectra of solvated amines in the THz range with pKa values. This demonstrates the potential of THz spectroscopy to probe the charge state of biomolecules in water in a label free manner.


Simulating the entropic collapse of coarse-grained chromosomes.
Depletion forces play a role in the compaction and decompaction of chromosomal material in simple cells, but it has remained debatable whether they are sufficient to account for chromosomal collapse. We present coarse-grained molecular dynamics simulations, which reveal that depletion-induced attraction is sufficient to cause the collapse of a flexible chain of large structural monomers immersed in a bath of smaller depletants. These simulations use an explicit coarse-grained computational model that treats both the supercoiled DNA structural monomers and the smaller protein crowding agents as combinatorial, truncated Lennard-Jones spheres. By presenting a simple theoretical model, we quantitatively cast the action of depletants on supercoiled bacterial DNA as an effective solvent quality. The rapid collapse of the simulated flexible chromosome at the predicted volume fraction of depletants is a continuous phase transition. Additional physical effects to such simple chromosome models, such as enthalpic interactions between structural monomers or chain rigidity, are required if the collapse is to be a first-order phase transition.


[HLA-E molecule induction on the surface of tumor cells protects them from cytotoxic lymphocytes].
Modern immunotherapy has developed powerful tools for mounting antitumor response which nevertheless have had only limited success in clinic. Tumor cells use different mechanisms to escape from immune system. Thus, one of the reasons of unsuccessful immunotherapy might be induction of tolerance of tumor-specific cytotoxic lymphocytes by tumor cells. Previously we have demonstrated expression of HLA-E molecule by the cells of melanoma cell lines. In this paper we have studied HLA-E-dependent mechanism of melanoma cell escape from immune response.


Reappraisal of Interpersonal Criticism in Social Anxiety Disorder: A Brain Network Hierarchy Perspective.
Emotion regulation is thought to involve communication between and within large-scale brain networks that underlie emotion reactivity and cognitive control. Aberrant network interaction might therefore be a key neural feature of mental disorders that involve emotion dysregulation. Here we tested whether connectivity hierarchies within and between emotion reactivity and cognitive reappraisal networks distinguishes social anxiety disorder (SAD) patients (n = 70) from healthy controls (HC) (n = 25). To investigate network organization, we implemented a graph-theory method called Dependency Network Analysis. Participants underwent fMRI while watching or reappraising video clips involving interpersonal verbal criticism. During reappraisal, the reappraisal network exerted less influence on the reactivity network in SAD participants. Specifically, the influence of the right inferior frontal gyrus on both reappraisal and reactivity networks was significantly reduced in SAD compared with HC, and correlated negatively with negative emotion ratings among SAD participants. Surprisingly, the amygdala exhibited reduced influence on the reappraisal network in SAD relative to HC. Yet, during the watch condition, the left amygdala's influence on the reactivity network increased with greater social anxiety symptoms among SAD participants. These findings refine our understanding of network organization that contributes to efficient reappraisal or to disturbances in applying this strategy in SAD.


HD-tDCS induced changes in resting-state functional connectivity: Insights from EF modeling.
High-definition transcranial direct current stimulation (HD-tDCS) holds promise for therapeutic use in psychiatric disorders. One obstacle for the implementation into clinical practice is response variability. One way to tackle this obstacle is the use of Individualized head models. This study investigated the variability of HD-tDCS induced electric fields (EFs) and its impact on resting-state functional connectivity (rsFC) during different time windows. In this randomized, double-blind, and sham controlled study, seventy healthy males underwent 20 min of 1.5 mA HD-tDCS on the right inferior frontal gyrus (rIFG) while undergoing resting-state functional magnetic resonance imaging (rs-fMRI). Individual head models and EF simulations were created from anatomical images. The effects of HD-tDCS on rsFC were assessed using a seed-to-voxel analysis. A subgroup analysis explored the relationship between EF magnitude and rsFC during different stimulation time windows. Results highlighted significant variability in HD-tDCS-induced EFs. Compared to the sham group, the active group showed increased rsFC between the rIFG and the left prefrontal cortex, during and after stimulation. During active stimulation, EF magnitude correlated positively with rsFC between the rIFG and the left hippocampus initially, and negatively during the subsequent period. This study indicated an HD-tDCS induced increase of rsFC between left and right prefrontal areas. Furthermore, an interaction between the magnitude and the duration of HD-tDCS on rsFC was observed. Due to the high EF variability that was apparent, these findings highlight the need for individualized HD-tDCS protocols and the creation of head models to optimize effects and reduce response heterogeneity.


Comparing growth velocity of HIV exposed and non-exposed infants: An observational study of infants enrolled in a randomized control trial in Zambia.
Impaired growth among infants remains one of the leading nutrition problems globally. In this study, we aimed to compare the growth trajectory rate and evaluate growth trajectory characteristics among children, who are HIV exposed uninfected (HEU) and HIV unexposed uninfected (HUU), under two years in Zambia. Our study used data from the ROVAS II study (PACTR201804003096919), an open-label randomized control trial of two verses three doses of live, attenuated, oral RotarixTM administered 6 &10 weeks or at 6 &10 weeks plus an additional dose at 9 months of age, conducted at George clinic in Lusaka, Zambia. Anthropometric measurements (height and weight) were collected on all scheduled and unscheduled visits. We defined linear growth velocity as the rate of change in height and estimated linear growth velocity as the first derivative of the mixed effect model with fractional polynomial transformations and, thereafter, used the second derivative test to determine the peak height and age at peak heigh. We included 212 infants in this study with median age 6 (IQR: 6-6) weeks of age. Of these 97 (45.3%) were female, 35 (16.4%) were stunted, and 59 (27.6%) were exposed to HIV at baseline. Growth velocity was consistently below the 3rd percentile of the WHO linear growth standard for HEU and HUU children. The peak height and age at peak height among HEU children were 74.7 cm (95% CI = 73.9-75.5) and 15.5 months (95% CI = 14.7-16.3) respectively and those for HUU were 73 cm (95% CI = 72.1-74.0) and 15.6 months (95% CI = 14.5-16.6) respectively. We found no difference in growth trajectories between infants who are HEU and HUU. However, the data suggests that poor linear growth is universal and profound in this cohort and may have already occurred in utero.


Neuroadaptive changes in mesocorticolimbic dopamine and acetylcholine neurons following cocaine or saline self-administration are dependent on pre-existing individual differences.
Previously, we demonstrated that stress-induced self-grooming behaviour in rats predicted an enhanced motivation to self-administer cocaine as determined under a progressive ratio schedule of reinforcement. The enhanced motivation of high grooming (HG) rats was associated with a reduced reactivity of dopaminergic neurons in the medial prefrontal cortex and amygdala, but not nucleus accumbens. In the present study, we studied the effect of cocaine and saline self-administration on these pre-existing differences in neurochemical profile by determining the electrically evoked release of [3H]dopamine and [14C]acetylcholine from superfused slices of the nucleus accumbens shell and core, medial prefrontal cortex and amygdala of HG and low grooming (LG) rats. Although HG and LG rats did not differ in acquisition of cocaine and saline self-administration, both conditions induced substantially different neuroadaptations in these rats. Differences in depolarisation-induced dopamine and acetylcholine release were maintained in the medial prefrontal cortex, emerged in the nucleus accumbens and dissipated in the amygdala. These results indicate that altered reactivity of mesocorticolimbic dopaminergic and cholinergic neurons due to exposure to cocaine and environmental stimuli (saline) is dependent on pre-existing neurochemical differences and displays region-specificity. These pre-existing differences and the cocaine- and environmental-induced neuroadaptations seem to act in concert to produce an enhanced motivational state to self-administer cocaine.


Tracking Mouse Trajectories Related to Decisions About Sexual Interest.
The current research explored continuous behavioral processes by recording participants' hand movements toward a response option using Mousetracker software. Across three studies, we tested the idea that sexual interest-related decisions can be indexed using automatic movements. Using a UK sample of self-identified same- and opposite-gender attracted men (N = 43), Study 1 showed that the process of approaching the response button was faster, more direct, and less curved when participants responded from their own (vs. a non-preferred) perspective. Study 2 supported these findings using a German sample of self-identified same- and opposite-gender attracted men (N = 66). Using a teleiophilic sample of 100 self-identified opposite-gender attracted men (n = 51) and women (n = 49), Study 3 examined age-specific sexual interests (i.e., responding to stimuli from a pedophilic and a teleiophilic perspective). As expected, in the teleiophilic block, participants' responses were faster, more direct, and less curved than in the pedophilic block. Collectively, the results suggest that mousetracking shows promise as a measure of sexual interest (across the dimension of both gender and age), providing grounds for future research.


Neuropeptide and neurohormone precursors in the pea aphid, Acyrthosiphon pisum.
Aphids respond to environmental changes by developing alternative phenotypes with differing reproductive modes. Parthenogenetic reproduction occurs in spring and summer, whereas decreasing day lengths in autumn provoke the production of sexual forms. Changing environmental signals are relayed by brain neuroendocrine signals to the ovarioles. We combined bioinformatic analyses with brain peptidomics and cDNA analyses to establish a catalogue of pea aphid neuropeptides and neurohormones. 42 genes encoding neuropeptides and neurohormones were identified, of which several were supported by expressed sequence tags and/or peptide mass analyses. Interesting features of the pea aphid peptidome are the absence of genes coding for corazonin, vasopressin and sulfakinin and the presence of 10 different genes coding insulin related peptides, one of which appears to be very abundantly expressed.


Susceptibility to Heart Defects in Down Syndrome Is Associated with Single Nucleotide Polymorphisms in HAS 21 Interferon Receptor Cluster and VEGFA Genes.
Congenital heart defects (CHDs) are present in about 40-60% of newborns with Down syndrome (DS). Patients with DS can also develop acquired cardiac disorders. Mouse models suggest that a critical 3.7 Mb region located on human chromosome 21 (HSA21) could explain the association with CHDs. This region includes a cluster of genes (IFNAR1, IFNAR2, IFNGR2, IL10RB) encoding for interferon receptors (IFN-Rs). Other genes located on different chromosomes, such as the vascular endothelial growth factor A (VEGFA), have been shown to be involved in cardiac defects. So, we investigated the association between single nucleotide polymorphisms (SNPs) in IFNAR2, IFNGR2, IL10RB and VEGFA genes, and the presence of CHDs or acquired cardiac defects in patients with DS. Individuals (n = 102) with DS, and age- and gender-matched controls (n = 96), were genotyped for four SNPs (rs2229207, rs2834213, rs2834167 and rs3025039) using KASPar assays. We found that the IFNGR2 rs2834213 G homozygous genotype and IL10RB rs2834167G-positive genotypes were more common in patients with DSand significantly associated with heart disorders, while VEGFA rs3025039T-positive genotypes (T/*) were less prevalent in patients with CHDs. We identified some candidate risk SNPs for CHDs and acquired heart defects in DS. Our data suggest that a complex architecture of risk alleles with interplay effects may contribute to the high variability of DS phenotypes.


Energetic sub-2-cycle laser with 216 W average power.
Few-cycle lasers are essential for many research areas such as attosecond physics that promise to address fundamental questions in science and technology. Therefore, further advancements are connected to significant progress in the underlying laser technology. Here, two-stage nonlinear compression of a 660 W femtosecond fiber laser system is utilized to achieve unprecedented average power levels of energetic ultrashort or even few-cycle laser pulses. In a first compression step, 408 W, 320 μJ, 30 fs pulses are achieved, which can be further compressed to 216 W, 170 μJ, 6.3 fs pulses in a second compression stage. To the best of our knowledge, this is the highest average power few-cycle laser system presented so far. It is expected to significantly advance the fields of high harmonic generation and attosecond science.


The metabolism of lipids in mouse pancreatic islets. The oxidation of fatty acids and ketone bodies.
The rate of oxidation of 14C-labelled fatty acids and of ketone bodies was measured in isolated pancreatic islets of obese-hyperglycaemic mice (ob/ob). The following main observations were made. 1. Octanoate, palmitate and oleate were all converted into CO2 by the pancreatic islets. Octanoate was oxidized with the highest rate followed by palmitate and oleate. 2. The rate of oxidation of 0.7 mM-palmitate was 3.1 pmol/h per mug drug weight. This was decreased by 50% in the presence of 16.7 mM-glucose. The rate of palmitate oxidation was also inhibited by 2-bromostearate. The palmitate oxidation showed a concentration-dependent increase, which was most marked between 0.25 and 1.0 mM. 3. Octanoate (5 mM) had no effect on the rate of oxidation of 3.3 mM- glucose. 4. Acetoacetate (5 mM) and D-3-hydroxybutyrate (5 mM) were oxidized at rates of 5.9 and 5.4 pmol/h per mug dry weight respectively. These rates were less than 10% of those found in kidney-cortex slices. The magnitude of the oxidation rates found for fatty acids and for ketone bodies suggest that these substrates represent important metabolic fuels for the pancreatic B-cells.


Ovulation induction with gonadotropins causes increased sister chromatid exchanges.
Gonadotropins are widely accepted agents for ovulation induction in infertile women. On the other hand, several authors discuss the possible effect of gonadotropins on the developmental mechanism of ovarian cancer. SCE is a method of genotoxicity investigation and it is an excellent parameter to monitor the DNA damage and repair. There are numbers of studies showing the relationship between endogenous or exogenous hormones and SCEs. The aim of this study was to investigate with SCE techniques the effects of long-term (6 months) use of gonadotropins on DNA as we couldn't find any other study on the effect of long term use. We found increased sister chromatid exchange rates in a study group as compared to a control group. This may be one of the causes of increased ovarian cancer risk in infertile population.


Rituximab treatment did not aggravate ongoing progressive multifocal leukoencephalopathy in a patient with multiple sclerosis.
A multiple sclerosis (MS) patient developed progressive multifocal leukoencephalopathy (PML) after 43 months of natalizumab treatment. New clinical and magnetic resonance imaging (MRI) findings were initially misinterpreted as breakthrough MS disease activity and natalizumab treatment was replaced by rituximab treatment. The patient had a single infusion of rituximab 1000 mg before a definite PML diagnosis was confirmed. Despite undetectable levels of B-cells, JC virus DNA became undetectable in the cerebrospinal fluid by quantitative polymerase chain reaction. The patient partially recovered without any clinical or MRI signs of new MS activity. These findings suggest that B-cell depletion in a non-immune compromised individual did not prevent the patient from clearing the JC virus infection.


The effect of heparin and EDTA on the NBT test.
Heparin precipitates nitroblue tetrazolium (NBT) in the absence of protein (plasma). This NBT-heparin precipitate, when presented to neutrophils, results in NBT scores higher than those obtained with NBT in solution. It is postulated that the high NBT scores obtained with increasing heparin concentrations are due to increased NBT precipitation and hence increased phagocytosis. It is proposed that the variability of NBT scores obtained when heparin is used as the anticoagulant is related to the amount of precipitate formed and to inhibition of phagocytosis by excess heparin. EDTA, in the presence of plasma and NBT, forms a precipitate consisting of protein and little, if any, NBT. In the presence of EDTA, NBT scores are lower than those obtained with heparin. EDTA, at concentrations used to prevent coagulation, inhibits phagocytosis and this could explain the low NBT scores. These observations provide a mechanism for entry of NBT dye into neutrophils and may help explain the inconsistent results found in both normal and disease states.


Ocular screening adherence across Hispanic/Latino heritage groups with diabetes: results from the Ocular SOL ancillary to the Miami site of the Hispanic Community Health Study/Study of Latinos (HCHS/SOL).
The objective of this study was to examine the prevalence and correlates of ocular screening adherence among select Hispanics/Latinos living with diabetes. Data were obtained through an ancillary study of the Hispanic Community Health Study/Study of Latinos (Miami site). Participants included Hispanics/Latinos aged 40+ years who underwent a baseline examination/risk factor assessment (2008-2011) and then completed a survey on vision health/knowledge (conducted October 2011-September 2013; sample n=1235; diabetic subsample=264). The dependent variable was having a dilated eye examination within the past 12 months. Covariate candidate selection for entry into sequential multivariable logistic regression models was guided by Anderson's Behavioral Model of Health Services Use and the Behavioral Model for Vulnerable Populations. Participants aged 65+ were more likely to have dilated eye examinations (OR 2.62, 95% CI 1.22 to 5.60) compared with those aged 40-54 years. Participants less likely to have dilated examinations had a high school degree or general educational development (GED) (OR 0.30, 95% CI 0.10 to 0.96, compared to no degree) and those currently uninsured or never insured ((OR 0.34, 95% CI 0.14 to 0.83) and (OR 0.19, 95% CI 0.07 to 0.51)) compared to those currently insured. Participants who heard or saw something about eye health from two or more sources (eg, media outlets, doctor's office, relatives/friends) compared to those who reported no sources in the past 12 months were more likely to have a dilated eye examination (OR 2.82, 95% CI 1.26 to 6.28). Lack of health insurance is strongly associated with low screening uptake in Hispanics/Latinos living with diabetes. Health promotion strategies stressing the importance of annual dilated eye examinations and increasing sources of information on eye health are other potential strategies to increase screening uptake in Hispanics/Latinos.


Capillary refilling time in newborn babies: normal values.
To assess the normal values of capillary refilling time (CRT) in healthy newborn babies; to assess the effect of different nursery containers (incubator, radiant warmer, crib), phototherapy, birthweight, gestational age, size for gestational age and sex on CRT; to compare CRT at different body sites as well as to assess the variation between observers. Healthy neonates (n = 469) of different gestational ages and different sizes for gestational age, were studied 1 to 7 days after birth. CRT was measured in four of the most suitable sites-namely, midpoints of the sternum and the forehead, the palm of the hand and the plantar surface of the heel (defined as chest, head, palm and heel, respectively). The applied pressing time was 5 seconds. CRT was measured with a manual stopwatch. Only the chest and the head distribution curves followed the Gaussian curve. The mean values and standard deviation of CRT in all tested nursery containers, including phototherapy for the chest, ranged from 1.82 (0.34) seconds to 2.01 (0.423) seconds, and for the head from 1.59 (0.36) seconds to 1.83 (0.31) seconds. The mean value of chest CRT was always longer than the head CRT for all parameters. Significant differences were found between different nursery containers, receivers, and non-receivers of phototherapy and between observers. No difference was found between sex, birthweight, gestational age and size for gestational age. The upper limit of normal for neonatal CRT was 3 seconds. Nursery containers, phototherapy, and observers produced significantly different results, but the differences were not clinically important. CRT values of the midpoints of the sternum and the forehead are the most consistent.


Superagonistic CD28 Protects against Renal Ischemic Injury by Expansion of Regulatory T-Cell.
Regulatory T (Treg) cells are a highly suppressive subset of CD4+ lymphocytes and have recently been proved to be crucial to suppress the inflammatory responses of ischemic kidney injury. CD28 superagonists (CD28sa) are monoclonal antibodies that preferentially expand Treg cells without a T-cell receptor and a costimulatory signal. This study aims to test the protection and discover the mechanisms of CD28sa treatment against renal ischemia-reperfusion (IR) injury (IRI). Male C57BL/6N mice were treated with CD28sa via peritoneal injection (0.1 mg) 6 days before the induction of IRI, or with 18-min ischemic precondition (IPC). IRI was induced by bilateral clamping of renal pedicles for 35 min followed by reperfusion. The role of Treg expansion in renal protection conferred by CD28sa treatment was examined using anti-CD25 antibody. CD28sa treatment alone significantly increased the percentage of Treg cells in the spleen (18.10 ± 2.00 vs. 6.64 ± 0.86%, p < 0.01), peripheral blood (16.43 ± 5.94 vs. 2.57 ± 1.09%, p < 0.01), and kidney (2.69 ± 0.90 vs. 0.53 ± 0.14%, p < 0.01) of C57BL/6N mice 6 days after the administration. Mice pretreated with CD28sa or IPC had less renal injury at 24 h after IRI with attenuation of renal tubular damage and lower serum creatinine compared with the mice that underwent renal IRI alone. The number of infiltrating macrophages in the kidney and IFN-γ secreting CD4+ T cells in peripheral blood were diminished in the CD28sa-IR group and the IPC-IR group. The renal protection bestowed by CD28sa or IPC was abolished by anti-CD25 antibody administration. Treg expansion induced by CD28sa ameliorated renal IRI.


Complications of anterior cervical spine surgery: a systematic review of the literature.
The anterior approach to the cervical spine is commonly utilized for a variety of degenerative, traumatic, neoplastic, and infectious indications. While many potential complications overlap with those of the posterior approach, the distinct anatomy of the anterior neck also presents a unique set of hazards. We performed a systematic review of the literature to assess the etiology, presentation, natural history, and management of these complications. Following the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA), a PubMed search was conducted to evaluate clinical studies and case reports of patients who suffered a complication of anterior cervical spine surgery. The search specifically included articles concerning adult human subjects, written in the English language, and published from 1989 to 2019. The PubMed search yielded 240 articles meeting our criteria. The overall rates of complications were as follows: dysphagia 5.3%, esophageal perforation 0.2%, recurrent laryngeal nerve palsy 1.3%, infection 1.2%, adjacent segment disease 8.1%, pseudarthrosis 2.0%, graft or hardware failure 2.1%, cerebrospinal fluid leak 0.5%, hematoma 1.0%, Horner syndrome 0.4%, C5 palsy 3.0%, vertebral artery injury 0.4%, and new or worsening neurological deficit 0.5%. Morbidity rates in anterior cervical spine surgery are low. Nevertheless, the unique anatomy of the anterior neck presents a wide variety of potential complications involving vascular, aerodigestive, neural, and osseous structures.


OCT4 regulates WNT/β-catenin signaling and prevents mesoendoderm differentiation by repressing EOMES in porcine pluripotent stem cells.
The regulatory network between signaling pathways and transcription factors (TFs) is crucial for the maintenance of pluripotent stem cells. However, little is known about how the key TF OCT4 coordinates signaling pathways to regulate self-renewal and lineage differentiation of porcine pluripotent stem cells (pPSCs). Here, we explored the function of OCT4 in pPSCs by transcriptome and chromatin accessibility analysis. The TFs motif enrichment analysis revealed that, following OCT4 knockdown, the regions of increased chromatin accessibility were enriched with EOMES, GATA6, and FOXA1, indicating that pPSCs differentiated toward the mesoendoderm (ME) lineage. Besides, pPSCs rapidly differentiated into ME when the WNT/β-catenin inhibitor XAV939 was removed. However, the ME differentiation of pPSCs caused by OCT4 knockdown did not rely on the activation of WNT/β-catenin signaling because the target gene of WNT/β-catenin signaling, AXIN2 was not upregulated after OCT4 knockdown, despite significant upregulation of WLS and some WNT ligands. Importantly, OCT4 is directly bound to the promoter and enhancers of EOMES and repressed its transcription. Overexpression of EOMES was sufficient to induce ME differentiation in the presence of XAV939. These results demonstrate that OCT4 can regulate WNT/β-catenin signaling and prevent ME differentiation of pPSCs by repressing EOMES transcription.


[Clinical assessment of immune parameters at surgical patients with syndrome of systemic inflammatory reaction].
Results of immunological examination of 423 patients with syndrome of systemic inflammatory reaction of different ethiology (mediastinitis--78 patients, peritonitis--85, severe acute pancreatitis--91, trauma of thorax and abdomen complicated with massive hemorrhage--111, severe burn trauma--40 patients) are analyzed. For complex and objective assessment of immune reaction the system of scores has been developed. This scale permitted to detect the immune disorders, to diagnose the degree of immune insufficiency at early stages of disease, to predict the purulent complications and to start timely immune therapy.


Hearing Health in Agricultural Aviation Pilots from Cindacta II Wearing Earplugs and a Helmet.
Introduction Agricultural aviation pilots, exposed daily to intense vibration and noise, are likely to develop noise-induced hearing loss (NIHL). Objectives The aim of this study was to investigate the presence of audiograms consistent with NIHL in agricultural aviation pilots who use earplugs and helmets. Methods We conducted a cross-sectional cohort and observational study. The data were taken from the medical records and audiograms of 94 pilots. Results NIHL was identified in 9.5% of individuals with hearing loss by audiograms at 3,000, 4,000, or 6,000 Hz. Normal audiograms were observed in 46.8% of pilots surveyed. Bilateral hearing loss was more frequent than unilateral hearing loss, occurring in 64.8% of cases. Conclusion Although there was a low incidence of audiograms compatible with NIHL in the records of the pilots examined, the disorder still occurs despite the doubled use of individual hearing protection equipment (helmets and earplugs) for agricultural aviation pilots. Nevertheless, even with the use of earplugs and helmets as noise protectors, the data showed that agricultural pilots suffer inner ear damage caused by occupational noise. Prevention and periodic audiologic evaluations must be conducted in noise-exposed occupational groups.


Defective retinoic acid regulation of the Pit-1 gene enhancer: a novel mechanism of combined pituitary hormone deficiency.
Pit-1 is a pituitary-specific transcription factor responsible for pituitary development and hormone expression in mammals. Pit-1 contains two protein domains, termed POU-specific and POU-homeo, which are both necessary for DNA binding and activation of the GH and PRL genes and regulation of the PRL, TSH-beta subunit (TSH-beta), and Pit-1 genes. Pit-1 is also necessary for retinoic acid induction of its own gene during development through a Pit-1-dependent enhancer. Combined pituitary hormone deficiency is caused by defective transactivation of target genes in the anterior pituitary. In the present report, we provide in vivo evidence that retinoic acid induction of the Pit-1 gene can be impaired by a Pit-1 gene mutation, suggesting a new molecular mechanism for combined pituitary hormone deficiency in man.


Critical sarcomere extension required to recruit a decaying component of extra force during stretch in tetanic contractions of frog skeletal muscle fibers.
29 single frog skeletal muscle fibers were stretched during fused tetanic contractions. The force increase during stretch exhibited a breakpoint at a critical length change (average: 16.6 nm per one-half sarcomere) that was independent of velocity of stretch and of sarcomere length between 1.8 and 2.8 microns. After stretch there was an early decaying force component with a force-extension curve similar to that during stretch, which disappeared over approximately 2 s. This component was removed by a small, quick release, leaving a longer-lasting component. The critical amplitude of release required to produce this result was found by clamping the fiber to a load at which there was zero velocity of shortening. This amplitude increased with time up to the angle in the force record during stretch, was constant for the remainder of the stretch, and decreased with time after the end of stretch; it was consistently less than the critical amplitude of stretch required to reach the breakpoint of force enhancement during stretch but was also independent of sarcomere length. The force drop accompanying the critical release showed a small increase up to an optimum magnitude at 2.4--2.7 microns sarcomere length, with a decrease at longer lengths.


[Why cardiovascular rehabilitation services should include patients with intermittent claudication?].
Chronic peripheral arterial disease represents a frequent and underestimated localization of atherosclerosis and its management often appears to be inadequate. The association with ischemic heart disease, the weighty prevalence of coronary disease risk factors, the high cardiovascular rate of morbidity and mortality, the sharp reduction in the tenor of life and the well-being supervised physical training efficacy constitute the main reasons that transform Cardiac Rehabilitation into an ideal setting for the treatment of patients suffering from intermittent claudication. Thanks to the cultural patrimony of cardiologists, together with other professionals, such as psychologists, dietitians, physiotherapists and nurses, it is possible to initiate an multi-comprehensive treatment program. Besides, this type of management may decrease both morbidity and mortality as well as reduce symptoms and improve the patients' quality of life.


A 451-nucleotide circular RNA from cherry with hammerhead ribozymes in its strands of both polarities.
The sequence of 451 nucleotides of a cherry small circular RNA (csc RNA1) associated with a cherry disease has been determined. Both csc RNA1 and its complementary strand can form hammerhead structures similar to those found previously in other plant and animal small RNAs. In the branched secondary structure of lowest free energy of csc RNA1, the sequences involved in the hammerhead structures, which comprise approximately one-fourth of this RNA, are found opposite each other, forming part of a rod-like segment. Plus- and minus-strand full-length transcripts of csc RNA1 self-cleaved during transcription and after purification, as predicted by the hammerhead structures, which are stable and very probably act as single hammerhead structures. The minus-strand hammerhead structure of csc RNA1 is exceptional in having a central loop with only 11 conserved nucleotides, a situation previously observed in only one other natural hammerhead structure. Both hammerhead structures of csc RNA1 are also peculiar in having an A instead of a C preceding the self-cleavage sites. The in vivo concentration of the plus strand of csc RNA1 is only slightly higher than that of its complementary strand, and significant fractions of both strands are extracted from the tissue in the form of a complex. csc RNA1 has sequence similarities to viroids and especially to some viroid-like satellite RNAs; they also share some characteristics of their corresponding hammerhead structures with these satellite RNAs. These observations, together with the association in symptomatic tissue of csc RNA1 with a set of presumably viral double-stranded RNAs, suggest that csc RNA1 is a new viroid-like satellite RNA.


Formulation of Saudi Propolis into Biodegradable Chitosan Chips for Vital Pulpotomy.
Propolis has been widely used to treat oral cavity disorders, such as endodontal and periodontal diseases and microbial infections. The study aimed at the formulation of commercial Saudi propolis into biodegradable chitosan chips and evaluation of its effectiveness as a pulpotomy agent. The standardization of 80% ethanolic propolis extract was performed regarding its total phenolic content, total flavonoid content, quantitative estimation of main polyphenolic constituents and antioxidant activity. Chitosan chips containing propolis extract were prepared by the solvent/ casting method. The investigated variables were % of chitosan polymer (2, 2.5 and 3%), % of plasticizer (1, 5 and 10%) and incorporation of different concentrations of hydroxypropyl methylcellulose (5, 10 and 20% of polymer weight). The chips were characterized for weight and thickness uniformity, content uniformity, pH, percentage moisture loss, swelling index, tensile strength and in vitro propolis release. The optimal propolis chip formulation was further investigated in dogs regarding the short term response of primary dental pulp to propolis chips compared with the most commonly used formocresol preparation. The prepared films were flexible and demonstrated satisfactory physicochemical characteristics. The optimal formulation showed an initial release of about 41.7% of the loaded propolis followed by a sustained release extended up to 7 days. The kinetics study demonstrated that propolis release was controlled by Fick´s diffusion. The optimal propolis chip formulation resulted in less pulpal inflammation compared to formocresol, and produced hard tissue formation in all specimens. Formulation of commercial Saudi propolis as a biodegradable chitosan chip is an effective alternative to the commercially available chemical agents for the treatment of vital pulpotomy.


Antistaphylococcal activity of a cyclic peptide, LY146032, and vancomycin.
The inhibitory and bactericidal activities of a novel cyclic peptide antibiotic, LY146032, and vancomycin against oxacillin-susceptible and oxacillin-resistant staphylococci were compared. The MICs for 90% of strains were two- to fourfold higher for vancomycin than for LY146032. MBC/MIC ratios for all strains were less than or equal to 2. In killing rate studies with four strains of staphylococci, there was no detectable growth in the presence of 4X the MIC of either LY146032 or vancomycin after 24 h of incubation.


Inhibition of penetration of cultured cells by Eimeria bovis sporozoites by monoclonal immunoglobulin G antibodies against the parasite surface protein P20.
Five monoclonal antibodies (MAbs) were partially characterized and tested for their ability to inhibit penetration of Madin-Darby bovine kidney (MDBK) cells by sporozoites of Eimeria bovis. By indirect fluorescent-antibody assays, all MAbs reacted with acetone-fixed sporozoites, but only two MAbs, EbS9 (immunoglobulin G1) and EbS11 (immunoglobulin G2a), localized specifically on the plasmalemma of live sporozoites. Two of the five MAbs also reacted with acetone-fixed first-generation merozoites of E. bovis; however, none of the MAbs reacted with live merozoites. Treatment of live sporozoites with EbS9 or EbS11 resulted in 79 and 73% decreases, respectively, in sporozoite penetration of MDBK cells. No significant differences in cell penetration occurred in MDBK cells inoculated with sporozoites that had been treated with the other three MAbs. Both EbS9 and EbS11 reacted in Western blots (immunoblots) of sporozoites with the same 20,000-relative-molecular-weight protein. The antigens against which these neutralizing MAbs react might be useful in immunizing against bovine coccidiosis.


Effect of morphine analogues on chemotaxis in Escherichia coli.
Pretreatment of Escherichia coli w3110 with levorphanol, a morphine analogue, reduced chemotaxis to serine, aspartic acid and galactose. This decreased chemotaxis was not due to decreased viability or motility. Pretreatment with 1.1 mM-levorphanol for 1 h, followed by washing to remove the drug prior to determination of chemotaxis, inhibited chemotaxis to each of the attractants by at least 80%. Pretreatment with dextrorphan, the enantiomorph of levorphanol, or levallorphan, the N-allyl analogue of levorphanol, resulted in a similar inhibition of chemotaxis. Reversal of the inhibition produced by pretreatment with levorphanol required a period of growth of at least one generation time.


Bone marrow transplantation in a patient with chronic myeloid leukemia and achondroplasia.
A 25-year-old female with achondroplastic dwarfism, diagnosed when she was six months old, was diagnosed with chronic myeloid leukemia (CML) in chronic phase with 9;22 translocation, A2B3 gene rearrangement and a Sokal index of 0.5. A recombinant interferon-alpha 2A treatment was started in October '94, which obtained a hematological, but not a karyotypic, response at the 6th and 12th month. In August '94, the patient was submitted to allogeneic bone marrow transplantation (alloBMT) from a 36-year-old matched sibling. The procedure was well tolerated. Disease-free survival (DFS) and overall survival (OS) were 21 and 39 months, respectively. We have not found any reported cases of achondroplastic patients submitted to alloBMT in the literature or in the IBMT Registry.