Vitamin D receptor (VDR) gene polymorphism influences the risk of osteoporosis in postmenopausal women of Northwest India.
The influence of VDR gene for the risk of osteoporosis has remained inconclusive. VDR gene polymorphism in relation to BMD in postmenopausal women of Northwest India revealed a susceptibility haplotype AGT. Possession of this haplotype exacerbates the risk of osteoporosis by 2.8 times, which manifests in recessive mode of inheritance. The purpose of this study is to understand the influence of coordinated effect of various single nucleotide polymorphisms (SNPs) within vitamin D receptor (VDR) gene for the risk of osteoporosis, which has remained undefined so far. Four pertinent SNPs of VDR gene, i.e., rs2228570, rs1544410, rs17879735, and rs731236 were examined with polymerase chain reaction-restriction fragment length polymorphism in dual energy X-ray absorptiometry verified 188 osteoporotics, 115 osteopenics, and 147 normal postmenopausal women of Northwest India. Minor allele 'T' of rs2228570 showed significant influence for the risk of osteoporosis (OR 1.60, 95%CI 1.16-2.20, P=0.004) and also in dominant (OR 2.32, 95%CI 1.47-3.64, P=0.0006) and additive model (OR 2.41, 95%CI 1.49-3.87, P=0.0006) after Bonferroni correction. Minor allele (T) of rs2228570 showed an allele dose effect with BMD of L1-L4 (P=0.009) and FN (P=0.036). Disease association analysis exposed a susceptibility haplotype AGT which influences the risk of osteopenia (OR 2.04, 95%CI 1.03-4.08, P=0.036) and osteoporosis (OR 2.90, 95%CI 1.61-5.38, P=0.00005) after adjusting the effects of age, BMI and years since menopause. This haplotype is significantly associated with BMDs at lumbar spine (P=0.0001) and femoral neck (P=0.016). In-depth analysis of this haplotype with other methods of Wald statistics and Akaike information criterion confirmed that carriers of each unit of this haplotype AGT increases the risk of osteoporosis by a factor of 2.80±0.34 (β±SE) which manifests (P=0.1 × 10⁻⁶) in its recessive mode of inheritance.


The beta 1 and beta 2 subunits of the AP complexes are the clathrin coat assembly components.
The beta 1 and beta 2 subunits are the closely-related large chains of the trans-Golgi network AP-1 and the plasma membrane AP-2 clathrin-associated protein complexes, respectively. Recombinant beta 1 and beta 2 subunits have been generated in Escherichia coli. It was found that, in the absence of all the other AP subunits, beta 1 and beta 2 interact with clathrin and drive the efficient assembly of clathrin coats. In addition, beta 2 subunits and AP complexes compete for the same clathrin binding site. The appearance of the clathrin/beta coats is the same as the barrel-shaped structures formed with native AP complexes. It is proposed that the principal function of the beta subunits is to initiate coat formation, while the remaining subunits of the AP complexes have other roles in coated pit and coated vesicle function.


Structure of the host-recognition device of Staphylococcus aureus phage ϕ11.
Phages play key roles in the pathogenicity and adaptation of the human pathogen Staphylococcus aureus. However, little is known about the molecular recognition events that mediate phage adsorption to the surface of S. aureus. The lysogenic siphophage ϕ11 infects S. aureus SA113. It was shown previously that ϕ11 requires α- or β-N-acetylglucosamine (GlcNAc) moieties on cell wall teichoic acid (WTA) for adsorption. Gp45 was identified as the receptor binding protein (RBP) involved in this process and GlcNAc residues on WTA were found to be the key component of the ϕ11 receptor. Here we report the crystal structure of the RBP of ϕ11, which assembles into a large, multidomain homotrimer. Each monomer contains a five-bladed propeller domain with a cavity that could accommodate a GlcNAc moiety. An electron microscopy reconstruction of the ϕ11 host adhesion component, the baseplate, reveals that six RBP trimers are assembled around the baseplate core. The Gp45 and baseplate structures provide insights into the overall organization and molecular recognition process of the phage ϕ11 tail. This assembly is conserved among most glycan-recognizing Siphoviridae, and the RBP orientation would allow host adhesion and infection without an activation step.


The effect of aging on the mineral status of female mice.
To clarify the effect of aging on the mineral status of female mice, mineral concentrations in their tissues were determined. Five 2-mo-old, five 6-mo-old, and five 10-mo-old female B10BR mice were fed a commercial diet. Iron, zinc, copper, calcium, magnesium, sodium, and potassium concentrations in the blood, liver, kidney, heart, brain, lung, and spleen of the mice were determined using a flame atomic absorption spectrophotometer. Iron concentrations in the liver, kidney, heart, brain, and spleen increased with age. Significant differences were detected between mice 2 and 6 mo of age and between mice 2 and 10 mo of age. Zinc concentrations in the heart and lung decreased significantly with age. Zinc concentrations in the heart and lung of 10-mo-old mice were significantly lower than those of 2-mo-old mice. It is noteworthy that the copper concentration in the brain of 10-mo-old mice was markedly higher compared with that of younger mice. Calcium accumulation was apparent in the kidney of mice at 10 mo.


Secretion, surface localization, turnover, and steady state expression of protein disulfide isomerase in rat hepatocytes.
Protein disulfide isomerase in isolated rat hepatocytes was present at a concentration of 7 micrograms/mg cell protein, representing a approximately 2-fold enrichment compared to isolated hepatic non-parenchymal cells. Though localized mainly in microsomal fractions of hepatocytes, direct immunofluorescence and cell surface radioiodination followed by immunoprecipitation revealed the presence of M(r) 57,000 disulfide isomerase at the cell surface. Electrostatic interaction of the protein with the cell surface was suggested by susceptibility to carbonate washing. Metabolic radiolabeling and immunoprecipitation studies also indicated that some of the newly synthesized M(r) 57,000 disulfide isomerase was secreted. Treatment of cells with colchicine markedly reduced the recovery of disulfide isomerase from the media, indicating microtubular-directed secretion of the protein. Partial staphlococcal V8 proteolytic digestion of the secreted protein revealed a peptide pattern similar to that of the cellular protein. Immunoprecipitation with antibody specific to the -KDEL peptide retention sequence confirmed the presence of this sequence in the secreted protein. Studies of the turnover of disulfide isomerase revealed a half-life of approximately 96 h. Treatment of cells with tunicamycin or heat shock resulted in an increased recovery of newly synthesized disulfide isomerase from cell lysates but diminished recovery from the media. The secretion and cell surface distribution of disulfide isomerase in hepatocytes may be important for the pathogenesis of immune mediated liver injury.


The sternocleidomastoid tumor of infancy.
The sternocleidomastoid tumor of infancy is an uncommon clinical entity which has received little attention in the otolaryngological literature. The diagnosis must be considered in any infant presenting with a lateral neck mass. Although its natural history favors spontaneous regression, the mass may initially grow in size. The purpose of treatment is the prevention of torticollis and craniofacial asymmetry. The initial treatment is non-surgical, passive and active exercises. Only when this treatment fails should surgery be performed. The disappearance of the lesion is not always a predictable sign. A small number of children will develop delayed torticollis or craniofacial asymmetry. It is important that parents be informed of this possibility and of the necessity for long term follow-up.


Virulence and Ecology of Agrobacteria in the Context of Evolutionary Genomics.
Among plant-associated bacteria, agrobacteria occupy a special place. These bacteria are feared in the field as agricultural pathogens. They cause abnormal growth deformations and significant economic damage to a broad range of plant species. However, these bacteria are revered in the laboratory as models and tools. They are studied to discover and understand basic biological phenomena and used in fundamental plant research and biotechnology. Agrobacterial pathogenicity and capability for transformation are one and the same and rely on functions encoded largely on their oncogenic plasmids. Here, we synthesize a substantial body of elegant work that elucidated agrobacterial virulence mechanisms and described their ecology. We review findings in the context of the natural diversity that has been recently unveiled for agrobacteria and emphasize their genomics and plasmids. We also identify areas of research that can capitalize on recent findings to further transform our understanding of agrobacterial virulence and ecology.


Comparison and Investigation of Exosomes from Human Amniotic Fluid Stem Cells and Human Breast Milk in Alleviating Neonatal Necrotizing Enterocolitis.
In view of the devastating impact of neonatal necrotizing enterocolitis (NEC) on newborns, the research on its intervention is particularly important. Although exosomes from human amniotic fluid stem cells (AFSC) and human breast milk (HBM) can protect against NEC, their mechanisms remain unclear. Here, we intend to compare the intervention effects of two types of exosomes on NEC mouse model and reveal their respective regulatory mechanisms. In general, both AFSC-derived exosomes (AFSC-exos) and HBM-derived exosomes (HBM- exos) can alleviate NEC- associated intestinal injury, significantly reduce NEC score, and reduce systemic and ileal inflammation and NEC related brain injury during experimental NEC. However, the mode and mechanism of action of the two sources of exosomes were not identical. In vivo, the number of ileal crypts was more significantly restored after HBM-exos intervention than AFSC-exos, and in vitro, HBM-exos preferentially inhibited the inflammatory response of intestinal epithelial cells (IECs), whereas AFSC-exos preferentially regulated the migration of IECs. Mechanistically, GO and KEGG analyses revealed the different therapeutic mechanisms of AFSC-exos and HBM-exos in NEC. Taken together, our results illustrate that AFSC-exos and HBM-exos reduce the severity of experimental NEC and intestinal damage through different mechanisms, supporting the potential of cell-free or breast milk free exosome therapy for NEC.


The Use of Internet and Social Media for Health Information and Its Consequences Among the Population in Saudi Arabia.
The Internet is being increasingly used in our lives. Along with Internet use, social media sites are especially popular and are used by many people on a daily basis. Many studies were conducted to see the people's perception and their attitude towards the use of the Internet for health information. Such studies were also carried out in Saudi Arabia, but most have been limited to a specific target sample of the population. Our study aimed to assess the perception towards and use of different platforms and search engines to seek health information in Saudi Arabia. A cross-sectional study was conducted in Saudi Arabia during the year 2021. An Arabic online questionnaire using Google forms was sent to a randomized sample. Microsoft Excel 2016 was used for data entry, and statistical analysis was performed using IBM© SPSS© Statistics version 25 (IBM© Corp., Armonk, NY, USA). Among the 1363 participants, 56.2% were females and 43.8% were males with a mean age of 30.73 ± 12.3. The majority were living in Western region. The most used social media platforms were WhatsApp (91.5%), YouTube (84.6%), and Twitter (82.6%), respectively. The most common medical websites browsed were the Saudi Ministry of Health (67%) and the Food and Drug Administration (54.4%). Some 40.1% of the participants had a medical consultation online from a doctor, and most of them (67.8%) trusted the online physician. Finally, most of the participants (90.9%) thought that health information on the Internet or social media contributes to raising the level of health awareness among the general population. There was a significant relationship between educational level and gender and online health-seeking behavior (p > 0.05). The study reinforced that health information that is sought from the Internet and social media platforms has a great impact on the population, emphasizing the need for credible information sources and how to access them.


Efficacy and safety of Treamid in the rehabilitation of patients after COVID-19 pneumonia: a phase 2, randomized, double-blind, placebo-controlled trial.
Many patients who recovered from COVID are still suffering from pulmonary dysfunction that can be persistent even for months after infection. Therefore, treatment to prevent irreversible impairment of lung function is needed. Treamid (bisamide derivative of dicarboxylic acid, BDDA) was shown to have anti-inflammatory and antifibrotic effects in animal models of pulmonary fibrosis. This study was designed to assess the safety, tolerability, and efficacy of Treamid in the rehabilitation of patients after COVID pneumonia. The aim was to establish whether Treamid could be effective in ameliorating post-COVID sequelae. The phase 2, randomized, double-blind, placebo-controlled clinical trial was done at 8 medical centers in Russia. Patients with a diagnosis of COVID in the past medical history (with the first symptoms of COVID appear no earlier than 2 months before screening) and having fibrotic changes in the lungs, decreased lung function (percentage of predicted FVC and/or DLCO < 80%), and moderate or severe dyspnea according to mMRC scale were enrolled and randomly assigned in a 1:1 ratio (stratified by the initial degree of lung damage, age, and concomitant chronic diseases) by use of interactive responsive technology to peroral administration of Treamid 50 mg or placebo once a day for 4 weeks. The primary outcome was the proportion of patients who achieved clinically significant improvement in FVC and/or DLCO (defined as a relative increase in FVC of ≥ 10% or a relative increase in FVC in the range of ≥ 5 to < 10% plus a relative increase in DLCO of ≥ 15%) at week 4 compared with baseline. Secondary endpoints included changes from baseline in dyspnea scoring evaluated by the modified Borg and mMRC scales, pulmonary function (FEV1, FVC, FEV1/FVC ratio, DLCO, TLC, FRC), 6-min walk distance, the overall score of the KBILD questionnaire, and the proportion of patients with a reduction in the degree of lung damage assessed by CT scores. This trial was registered on ClinicalTrials.gov (Identifier: NCT04527354). The study was fully funded by PHARMENTERPRISES LLC. 12 out of 29 patients (41%) in Treamid group achieved clinically significant improvement in FVC and/or DLCO compared to 5 out of 30 patients (17%) in placebo group (p = 0.036). There was a significant decrease of dyspnea according to modified Borg scale observed in the Treamid group (- 0.9 ± 0.7 vs. - 0.4 ± 0.8, p = 0.018). No significant differences in the adverse events were noted. Exploratory analysis of the female population indicated superiority of Treamid over placebo by decreasing dyspnea and the extent of lung damage as well as increasing TLC. 4 weeks oral administration of 50 mg Treamid was associated with clinically significant improvement in the post-COVID patients, evident by an increase in FVC and/or DLCO as well as decreasing dyspnea. Treamid was well tolerated and can be safely administered to patients discharged after COVID. Treamid was more effective in women visible by superior improvement of COVID sequalae after 4 weeks treatment. Considering that female gender is a risk factor associated with the development of post-COVID symptoms, Treamid might offer a pharmacological treatment for long-term sequalae after COVID and supports further investigation in future clinical trials in post-COVID patients.


REIN: a tool at the service of the elderly patients
On the occasion of the 20th anniversary of the REIN (French Renal Epidemiology and Information Network), a summary work on the contributions of the national French ESKD register was carried out. On the issue of elderly CKD patients, the following key messages were retained. The population of elderly dialysis patients continues to grow. These patients have multiple comorbidities including pathologies specific to the elderly like undernutrition, walking and cognitive disorders. The data from the REIN registry has been the subject of numerous studies pertaining to this group showing heterogeneous prognosis and treatment, requiring to be better evaluated to target an individualised treatment. Decision support tools have been developed using the registry data to help nephrologists and patients in deciding whether to start replacement therapy and whether to refer elderly patients for assessment for a renal transplant. Finally, the risk-benefit evaluation of the different replacement therapies in this population should be monitored at the registry level and on a case-by-case basis in clinical practice.


Mapping the governance of human resources for health in Serbia.
This article maps the current governance of human resources for health (HRH) in relation to universal health coverage in Serbia since the health sector reforms in 2003. The study adapts the Global Health Workforce Alliance/World Health Organization four-dimensional framework of HRH in the context of governance for universal health coverage. A set of proxies was established for the availability, accessibility, acceptability and quality of HRH. Analysis of official HRH documentation from relevant institutions and reports were used to construct a governance profile of HRH for Serbia from the introduction of the reform in 2003 up to 2013. The results show that all Serbian districts (except Sremski) surpass the availability threshold of 59.4 skilled midwives, nurses and physicians per 10,000 inhabitants. District accessibility of health workforce greatly differed from the national average with variances from +26% to -34%. Analysis of national averages and patient load of general practitioners showed variances among districts by ± 21%, whilst hospital discharges per 100 inhabitants deviated between +52% and -45%. Pre-service and in-service education of health workforce is regulated and accredited. However, through its efforts to respond to population health needs Serbia lacks a single coordinating entity to take overall responsibility for effective and coordinated HRH planning, management and development within the broader landscape of health strategy development.


Platelet transfusion tolerization by liver membrane treatment.
A nontoxic murine model has been developed in which sensitization to allogeneic platelet transfusions in adults can be prevented. This model is based upon allogeneic liver membrane (LM) induction of specific humoral tolerance to major histocompatibility alloantigens. In normal mice of the C3H background, syngeneic and H-2-incompatible congeneic platelets had a t1/2 = 15 +/- 3 hr; for mice of the C57BL background, the comparable t1/2 was 33 +/- 6 hr. Platelets of C3H background transfused into C57BL background recipients, and vice versa, had t1/22 midway between, 24 +/- 3 and 24 +/- 2 hr, respectively. These data suggest genetically determined variation in normal platelet survival. In passively immunized C3H background mice, the t1/2 was decreased to 10 +/- 2 hr. In C57BL background mice actively immunized i.p. with allogeneic lymphoid cells, t1/2 was decreased to 18 +/- 4 hr. When allogeneic LM was given concomitantly with the allogeneic cells, however, sensitization to foreign H-2 antigens was blocked, and survival of both C3H and C57BL background allogeneic platelets remained normal. These data demonstrate that in this free cell allograft system LM treatment is a safe and effective method for preventing adult sensitization to allogeneic platelets.


Short-term transcutaneous vagus nerve stimulation increases pupil size but does not affect EEG alpha power: A replication of Sharon et al. (2021, Journal of Neuroscience).
Transcutaneous auricular vagus nerve stimulation (taVNS) has been tested as a potential treatment for pharmaco-resistant epilepsy and depression. Its clinical efficacy is thought to depend on taVNS-induced activation of the locus coeruleus and other neuromodulator systems. However, unlike for invasive VNS in rodents, there is little evidence for an effect of taVNS on noradrenergic activity. We attempted to replicate recently published findings by Sharon et al. (2021), showing that short bursts of taVNS transiently increased pupil size and decreased EEG alpha power, two correlates of central noradrenergic activity. Following the original study, we used a single-blind, sham-controlled, randomized cross-over design. Human volunteers (n = 29) received short-term (3.4 s) taVNS at the maximum level below the pain threshold, while we collected resting-state pupil-size and EEG data. To analyze the data, we used scripts provided by Sharon and colleagues. Consistent with Sharon et al. (2021), pupil dilation was significantly larger during taVNS than during sham stimulation (p = .009; Bayes factor supporting the difference = 7.45). However, we failed to replicate the effect of taVNS on EEG alpha power (p = .37); the data were four times more likely under the null hypothesis (BF10 = 0.28). Our findings support the effectiveness of short-term taVNS in inducing transient pupil dilation, a correlate of phasic noradrenergic activity. However, we failed to replicate the recent finding by Sharon et al. (2021) that taVNS attenuates EEG alpha activity. Overall, this study highlights the need for continued research on the neural mechanisms underlying taVNS efficacy and its potential as a treatment option for pharmaco-resistant conditions. It also highlights the need for direct replications of influential taVNS studies.


Identifying features of a mobile-based application for self-care of people living with T2DM.
Diabetes self-care requires support to empower patients to improve self-monitoring and maintain the necessary self-care behaviors. We aimed to identify features of a mobile-based application as a technology-based device for self-care of people living with T2DM. This study was conducted in two main phases in 2020. In the first phase, a literature review study was performed to identify the data elements and technical features of the T2DM self-care application. In the second phase, using the information obtained from the review of similar articles, a questionnaire was designed to validate identified requirements. The statistical population of the present study consisted of 22 endocrinologists and metabolic specialists. Identification of 55 data elements and technical features for mobile-based self-care application for people with T2DM, and according to the statistical population, 15data elements for demographic requirements, 16 data elements for clinical requirements, and 17 features for the technical capability of this app were selected. Blood sugar monitoring, exercise, nutrition, weight monitoring, and educational capabilities were the most highlighted technical features of the T2DM self-care application. Software designers can use these requirements to design a self-care app for people with type-2 diabetes that can help manage and improve patients' health status.


In silico design and validation of a highly degenerate primer pair: a systematic approach.
The techniques of amplifying genetic materials have enabled the extensive study of several biological activities outside the biological milieu of living systems. More recently, this approach has been extended to amplify population of genes, from evolutionarily related gene family for detection and evaluation of microbial consortial with several unique potentialities (e.g., enzymatic degradability). Conceivably, primer mixtures containing substitutions of different bases at specific sites (degenerate primers) have enabled the amplification of these genes in PCR reaction. However, the degenerate primer design problem (DPD) is a constraint to designing this kind of primer. To date, different algorithms now exist to solve various versions of DPD problem, many of which, only few addresses and satisfy the criteria to design primers that can extensively cover high through-put sequences while striking the balance between specificity and efficiency. The highly degenerate primer (HYDEN) design software program primarily addresses this variant of DPD problem termed "maximum coverage-degenerate primer design (MC-DPD)" and its heuristics have been substantiated for optimal efficiency from significant successes in PCR. In spite of the premium presented for designing degenerate primers, literature search has indicated relatively little use of its heuristics. This has been thought to result from the complexity of the program since it is run only by command-line, hence limiting its accessibility. To solve this problem, researchers have optionally considered the manual design of degenerate primers or design through software programs that provides accessibility through a graphical user interface (GUI). Realizing this, we have attempted in this study to provide a user-friendly approach for researchers with little or no background in bioinformatics to design degenerate primers using HYDEN RESULTS: Virtual Tests of our designed degenerate primer pair through in silico PCR substantiated the correspondence between efficiency and coverage with the target sequences as pre-defined by the initial HYDEN output, thereby validating the potentials of HYDEN to effectively solve the MC-DPD problem. Additionally, the designed primer-pair mechanistically amplified all sequences used as a positive control with no amplification observed in the negative controls. In this study, we provided a turnkey protocol to simplify the design of degenerate primers using the heuristics of the HYDEN software program.


Personality Change Profiles and Changes in Cognition Among Middle-Aged and Older Adults.
Research on the relationship between personality traits and cognitive abilities has primarily used cross-sectional designs and considered personality traits individually in relation to cognitive dimensions. This study (N = 2,652) examined the relationship between Big Five personality change profiles and change in cognitive factors, episodic memory and executive functioning. Latent profile analysis was used to capture patterns of change across the Big Five traits. Three profiles of personality change were defined: Decreasers, Maintainers, and Increasers. The Decreasers declined more in episodic memory compared to the Increasers and Maintainers. Also, the Decreasers declined more in executive functioning compared to the Increasers, but not the Maintainers. The findings advance our understanding of the links between patterns of personality change and cognitive aging.


Serum levels of high mobility group box-1 protein (HMGB1) and soluble receptors of advanced glycation end-products (RAGE) in depressed patients treated with electroconvulsive therapy.
High mobility group box-1 (HMGB1) is one of the damage-associated molecular patterns produced by stress and induces inflammatory responses mediated by receptors of advanced glycation end-products (RAGE) on the cell surface. Meanwhile, soluble RAGE (sRAGE) exhibits an anti-inflammatory effect by capturing HMGB1. Animal models have shown upregulation of HMGB1 and RAGE in the brain or blood, suggesting the involvement of these proteins in depression pathophysiology. However, there have been no reports using blood from depressed patients, nor ones focusing on HMGB1 and sRAGE changes associated with treatment and their relationship to depressive symptoms. Serum HMGB1 and sRAGE concentrations were measured by enzyme-linked immunosorbent assay in a group of patients with severe major depressive disorder (MDD) (11 males and 14 females) who required treatment with electroconvulsive therapy (ECT), and also in a group of 25 age- and gender-matched healthy subjects. HMGB1 and sRAGE concentrations were also measured before and after a course of ECT. Depressive symptoms were assessed using the Hamilton Rating Scale for Depression (HAMD). There was no significant difference in HMGB1 and sRAGE concentrations in the MDD group compared to healthy subjects. Although ECT significantly improved depressive symptoms, there was no significant change in HMGB1 and sRAGE concentrations before and after treatment. There was also no significant correlation between HMGB1 and sRAGE concentrations and the HAMD total score or subitem scores. There were no changes in HMGB1 and sRAGE in the peripheral blood of severely depressed patients, and concentrations had no relationship with symptoms or ECT.


Four-compartment model and validation of deuterium dilution technique to estimate fat-free mass in Mexican youth.
To validate the measurement of fat-free mass (FFM) with the deuterium oxide (D(2)O) dilution technique (2C) against the four-compartment (4C) model in Mexican children. This was designed as a cross-sectional, non-probabilistic study. Sixty subjects (30 male and 30 female) 6-14 y of age were recruited and completed the study during 5 mo. Total body water was measured using the D(2)O dilution technique and FFM was calculated using Fomon's (6-10 y) and Lohman's (11-14 y) hydration constants. Body composition using the 4C model was calculated with Lohman's equation. Group mean accuracy showed no differences in FFM determined by D(2)O dilution and the 4C model (1.24 kg, P > 0.4), by gender (2.1 kg, P > 0.2), or by method-by-gender interaction (P > 0.7). FFMs were 26.9 and 25.7 kg by the 4C and 2C models, respectively. The test for coincidence of slopes and intercepts between the 2C and 4C models and the line of identity were not different (P > 0.05). Precision by R(2) explained 98% of the variance (standard error of the estimate 1.2 kg). Bias for the difference in FFM was not significant (-1.27, 95% confidence interval -1.5 to -0.9) and no association between the mean of the differences and the magnitude of the measurements was found (P > 0.05). Mean bias was -1.27 kg for FFM (P > 0.05), and limits of agreement were -3.1 to 0.8 kg. The D(2)O dilution technique used with these hydration constants was accurate, precise, and free of bias in Mexican children and adolescents compared with the 4C model.


From the Spectroscopic Reassessment of Authentic Alkaloid Samples to the Molecular Networking-Guided Discovery of Criophylline-Related Analogues from Callichilia inaequalis.
The molecular network-guided exploration of the alkaloid extract of Callichilia inaequalis stems revealed a cluster attributed tentatively to dimeric monoterpene indole alkaloids of the rare criophylline subtype, initiating the dual study reported herein. A patrimonial-themed portion of this work was aimed at performing a spectroscopic reassessment of criophylline (1), a monoterpene bisindole alkaloid for which the nature of the inter-monomeric connectivity and configurational assignments have remained dubious. A targeted isolation of the entity annotated as criophylline (1) was undertaken to strengthen the available analytical evidence. An extensive set of spectroscopic data was acquired from the authentic sample of criophylline (1a) isolated earlier by Cavé and Bruneton. These spectroscopic studies proved the samples to be identical, and the complete structure of criophylline could be assigned, half a century after it was first isolated. The absolute configuration of andrangine (2) was also ascertained based on a TDDFT-ECD approach from the authentic sample. The forward-looking aspect of this investigation resulted in the characterization of two new criophylline derivatives from C. inaequalis stems, namely, 14'-hydroxycriophylline (3) and 14'-O-sulfocriophylline (4). Their structures, including absolute configurations, were elucidated by analysis of NMR and MS spectroscopic data and by ECD analysis. Notably, 14'-O-sulfocriophylline (4) is the first sulfated monoterpene indole alkaloid to have been reported. The antiplasmodial activity against the chloroquine-resistant strain of Plasmodium falciparum FcB1 was determined for criophylline and its two new analogues.


Bacterial interactions and transport in unsaturated porous media.
The convection-dispersion transport model, which can well define solute transport, has been introduced to describe bacterial transport. Due to different interaction natures within the porous media, bacterial transport in the subsurface, especially in the vadose zone is a complex scenario. When transported in the vadose zone, bacteria may be captured on the media surface, at the air-water interface, or at the media-air-water three-phase interface depending upon the predominant interactions of concerned bacteria within the pore system. In this study, transport of Echerichia coli, Pseudomonas fluorescens and Bacillus subtilis in silica sand under water unsaturated conditions was investigated using column experiments. Bacterial interactions within the system were characterized based on bacterial and media surface thermodynamic properties, which were determined independently by means of contact angle measurements. These calculated interactions provided solid evidence of the bacterial retention mechanisms in the pore system, which served as the bases for suitable assumptions of bacterial transport modeling. The micro-scale interaction investigations helped eliminate uncertainties arising with bacterial transport modeling.


Stylissamide X, a new proline-rich cyclic octapeptide as an inhibitor of cell migration, from an Indonesian marine sponge of Stylissa sp.
A new proline-rich cyclic octapeptide named stylissamide X (1) was isolated from an Indonesian marine sponge of Stylissa sp. as an inhibitor of cell migration from the guidance of wound-healing assay. The chemical structure of stylissamide X (1) was determined on the basis of spectroscopic analysis, and stereostructure of the amino acids were deduced by Marfey's method. Compound 1 showed inhibitory activity against migration of HeLa cells in the ranges of 0.1-10 μM concentration through both wound-healing assay and chemotaxicell chamber assay, while the cell viability was maintained more than 75% up to 10 μM concentration of 1.


Extracapsular cataract extraction and posterior chamber intraocular lens implantation combined with trabeculectomy in patients with glaucoma.
We reviewed 75 consecutive cases of extracapsular cataract extraction and posterior chamber intraocular lens implantation combined with trabeculectomy in 69 patients with glaucoma. The mean preoperative intraocular pressure was 19.3 mm Hg on an average of 2.3 glaucoma medications. Visual acuity improved in 58 eyes (77%) at two months, with an average improvement of 3.3 and 3.6 lines at two and 12 months, respectively. Of the 75 eyes, 49 (65%) achieved a visual acuity of 20/40 or better; three patients (4%) had further deterioration in vision at the completion of follow-up because of progressive glaucoma or macular disease. Postoperatively, the average intraocular pressure was 3.8 and 3.0 mm Hg lower than the preoperative level at two and 12 months (P less than .001) on 0.63 and 0.79 glaucoma medications, respectively. However, 27 (36%) of the 75 eyes had a recorded intraocular pressure greater than 30 mm Hg and 30 (40%) had a pressure 7 mm Hg or more above their preoperative level during the first six months after surgery. Despite improved long-term control of intraocular pressure, detectable conjunctival filtering blebs were present in only 31 (41%) of 75 eyes at two months and in seven (12%) of 56 eyes at 12 months. Hyphema occurred in 34 (45%) of the cases.


The administration of beta carotene to prevent and regress oral carcinoma in the hamster cheek pouch and the associated enhancement of the immune response.
In the past four years this laboratory has utilized the hamster cheek pouch tumor model to investigate the anticancer activities of antioxidants, such as beta carotene. These molecules, which have exhibited no evidence of toxicity, have been administered systemically (oral ingestion), and locally to the tumor site in the hamster cheek pouch. The results have been either the inhibition of tumor growth, or the regression of tumor. Adjacent to the degenerating tumors a dense inflammatory infiltrate was observed. Specifically, the cytokines, tumor necrosis factor alpha, and beta, have been immunohistochemically localized to the site of regressed oral carcinoma. Recently, liposomes composed of phosphaditylcholine, phosphaditylserine, and phosphodityelanolamine were combined with beta carotene and injected locally to oral squamous cell carcinoma of the hamster. The results indicated that tumor cells accumulated the liposomes and were lysed while normal mucosal cells did not demonstrate this effect. Therefore antioxidants such as beta carotene can be localized to a tumor site, without a toxic response. Future studies on the anticancer activity of the antioxidants need to focus on the cellular and molecular changes produced in the immune effectors and in the mucosal cells following administration of the antioxidants.


Vacuum differential mobility spectrometry combined with column-switching liquid chromatography- mass spectrometry for the analysis of pyrrolizidine alkaloids in tea samples.
The benefit of combining liquid chromatography (LC), supercritical fluid chromatography (SFC) and vacuum Differential Mobility Spectrometry - Mass Spectrometry (vDMS-MS) was investigated for the analysis of fourteen diastereomeric pyrrolizidine alkaloids (PA); intermedine, echinatine, lycopsamine, indicine, intermedine-N-oxide, echinatine-N-oxide, indicine-N-oxide, lycopsamine-N-oxide, senecivernine, senecionine, jacobine, senecivernine-N-oxide, senecionine-N-oxide, retrorsine. The mobile phase composition (15-100% MeOH and ACN), flow rate (8-100 µL/min), vDMS cell pressure, and F value showed an effect on the mobility behavior of the analytes. At 15% MeOH with a flow rate of 100 µL/min and 33 mbar vDMS pressure, 8 out 14 PA could be partially or totally separated by vDMS-MS. As well as providing an additional separation dimension vDMS improved the selectivity and a 5-minute assay method was developed for the quantification of 10 out of 14 single diastereomeric PA in tea samples, using a short LC column-switching and hyphenated to vDMS-MS in the selected ion monitoring mode. The performance of the method was found to be comparable with a 12-minute standard LC-MS/MS method using detection in the selected reaction monitoring mode. Additionally, the combination of vDMS and SFC-MS was investigated and suggests that the mixture of CO2/MeOH influences the CV shifting of the PA to more negative compensation voltage, and the signal-to-noise ratio is improved by a factor of three compared to SFC-MS without vDMS.


Bacterial loads and antibiotic resistance profile of bacteria isolated from the most popular street food (Phuchka) in Bangladesh.
Phuchka is one of the most common street foods in Bangladesh. It is served with salad, sweet and sour tamarind dispersed water, and minced eggs as topping at places where people usually gather. This makes these foods susceptible to bacterial contamination. Therefore, assessing the bacterial load and antimicrobial profile of organisms isolated from phuchka and other foodstuffs served with it was the focus of this study. Bacterial isolates were isolated and identified from the samples after the bacterial loads were assessed as total viable count (TVC), total coliform count (TCC), and total staphylococcal count (TSC). The antibiotic resistance profile of the isolates was obtained using the disk diffusion method. Molecular detection of Escherichia coli isolates and the presence of gene responsible for tetracycline resistance was confirmed by polymerase chain reaction. According to the recommendations, the TVC value of 70% phuchka and egg samples was safe, whereas TSC value illustrated that 80% of both phuchka and egg samples were at safety level. For the TCC value, 80% egg and 70% phuchka samples were found to be safe for consumption. Among all the samples, the microbial loads of the vendors' hand wash were least in the safety level. Antibiotic sensitivity tests revealed that both Staphylococcus spp. and E. coli isolates were sensitive to gentamicin and ciprofloxacin but showed resistance to ampicillin. The data of this study indicate that phuchka can pose a public health problem as foodborne bacterial isolates which are antibiotic-resistant are found in it.


Effects of vagotomy on cholecystokinin- and dexfenfluramine-induced Fos-like immunoreactivity in the rat brain.
This study compared the effects of bilateral subdiaphragmatic vagotomy on the Fos-like immunoreactivity (FLI), a marker of neuronal activation, in rat brain induced by two anorectic agents, cholecystokinin (CCK) and the serotonin agonist, dexfenfluramine (DFEN). In the nonvagotomized rats, both CCK (5 micrograms/kg, IP) and DFEN (2 mg/kg, IP) induced FLI in the nucleus of the solitary tract (NST), the external subdivision of the lateral parabrachial nuclei (LPBE), the lateral subdivision of the central amygdaloid nucleus (CeL), and the bed nucleus of the stria terminalis (BST). However, subregional distribution of the FLI induced by the two agents was different in most of these regions. Additionally, the area postrema and the medial subdivision of the hypothalamic paraventricular nucleus were preferentially activated by CCK but not DFEN, while the caudate-putamen was activated by DFEN but not CCK. Bilateral subdiaphragmatic vagotomy completely abolished CCK-induced FLI in all the brain regions but did not attenuate DFEN-induced FLI in any of these regions, including the NST. The results of the present study suggest that DFEN-activation of the NST-LPBE-CeL/BST neuraxis is not mediated by the vagus nerve. On the other hand, and consistent with a variety of other data, activation of various parts of the brain by peripherally administered CCK depends on a vagal pathway. These data are discussed in relation to a previously proposed interaction between CCK and serotonin in mediating satiety.


Comparison of calcium effect on in vitro calcitonin and parathyroid hormone release by young and aged thyroparathyroid glands.
Serum immunoreactive parathyroid hormone (iPTH) and calcitonin (iCT) levels are higher in young than aged rats. However, serum calcium concentration does not change with age suggesting that the calcium regulation of PTH and CT secretion may be affected by aging. We compared iPTH and iCT secretion in vitro at low and high calcium concentrations using thyroparathyroid glands removed from young (2-3 months), adult (12-13 months), and old (24-27 months) F-344 male rats fed regular rat chow. Glands from each animal were incubated for 3 h in serum-free culture media containing 1.0 mM calcium and then transferred to media containing 2.5 mM calcium for another 3 h. Immunoreactive PTH and iCT concentrations of the media after each incubation period were determined by radioimmunoassay. Immunoreactive PTH and iCT secretion per pair of glands was significantly higher in glands from older animals regardless of calcium concentration. The decrease in iPTH, and increment in iCT, secretion in response to 2.5 mM calcium by glands from old rats was smaller than that observed for glands from young animals. These age-related changes in the regulation of secretion by calcium may contribute to the increased iPTH and iCT secretion and serum levels seen in older animals.


Air pollution and recurrence of cardiovascular events after ST-segment elevation myocardial infarction.
The effects of air pollution on discharged patients after ST-segment elevation myocardial infarction (STEMI) still remain uncertain. We examined the association between air pollutants and recurrent cardiovascular events in STEMI survivors. A retrospective cohort of 1641 discharged patients after STEMI was established in 2013 and followed until the end of 2019. Concentrations of air pollutants including fine particles <2.5 μm aerodynamic diameter (PM2.5), inhalable particles <10 μm aerodynamic diameter (PM10), sulfur dioxide (SO2), nitrogen dioxide (NO2), carbon monoxide (CO) and ozone (O3) measured by fixed ambient air monitoring stations were collected for exposure assessment. Multivariate-adjusted Cox proportional hazards models were used to estimate the increased risks of recurrent cardiovascular events. Compared with the first exposure quartile, for short-term exposure, hazard ratios (HRs) of recurrent cardiovascular events associated with the fourth exposure quartiles of PM2.5, PM10, NO2, SO2, CO, and O3 were 4.06 (95% CI: 2.62-6.30), 3.79 (95% CI: 2.57-5.58), 2.22 (95% CI: 1.67-2.94), 4.47 (95% CI: 3.08-6.48), 3.73 (95% CI: 2.54-5.48), and 5.35 (95% CI: 3.12-9.20), respectively. For long-term exposure, HRs associated with the fourth exposure quartiles of PM2.5, PM10, NO2, SO2, CO, and O3 were 6.43 (95% CI: 3.60-11.47), 4.77 (95% CI: 2.85-7.99), 3.22 (95% CI: 2.00-5.19), 3.20 (95% CI: 2.05-5.01), 4.44 (95% CI: 2.65-7.45), and 1.07 (95% CI: 0.80-1.42), respectively. The risks of recurrent cardiovascular events brought by air pollutants mostly increased nonlinearly. Short- and long-term exposure to air pollutants except ozone increases the risks of recurrent cardiovascular events in STEMI survivors. Better environmental policies and secondary prevention strategies should be developed to protect STEMI survivors as a susceptible population.


Characterization of RNA aptamers directed against the nucleocapsid protein of Rift Valley fever virus.
Nucleocapsid protein (N) is an essential RNA binding protein in many RNA viruses. During replication, N protein encapsidates viral genomic and antigenomic RNA, but not viral mRNA or other cellular RNAs. To discriminate between different species of RNA in a host cell, it is likely that N interacts with specific sequences and/or secondary structures on its target RNA. In this study, we explore the RNA binding properties of N using both natural and artificially selected RNAs as ligands. We found that N binds to RNAs that resemble the terminal panhandle structures of RVFV genomic and antigenomic RNA. Furthermore, we used SELEX to isolate RNA aptamers that bound N with high affinity and determined that N specifically recognizes and binds to GAUU and pyrimidine/guanine motifs. Interestingly, BLAST analysis revealed the presence of these motifs within the coding region of the viral genome, suggesting that N may interact with non-terminal viral RNA sequences during replication. Finally, the aptamer RNAs were used to construct a sensitive fluorescence based sensor of N binding with potential applications for drug screening and imaging methodologies.


The relative prognostic significance of total cathepsin D and HER-2/neu oncogene amplification in breast cancer. The South Australian Breast Cancer Study Group.
Total tumor cathepsin D (TCD) levels were determined prospectively by a radioimmunometric assay in tumor cytosol of 858 primary breast cancer patients diagnosed between 1989-1991. In 581 of these patients, tumor HER-2/neu oncogene amplification was simultaneously determined. In a "training-set" of 313 patients, "high" TCD was associated with significantly shorter disease-free survival (DFS). For the whole group, there was no correlation between TCD and pathologic stage, number of axillary nodes with tumor deposits, tumor size, histologic type and grade, or hormone receptor levels. In the node-positive group, high TCD level was associated with HER-2/neu amplification. After a median follow-up duration of 31 months, univariate analysis indicated that high TCD level was significantly associated with shorter DFS only in node-positive patients. The shorter DFS in association with high TCD levels was observed in both estrogen-receptor-positive and -negative patients. Cox multivariate analysis of DFS confirmed that high TCD level was predictive of shorter DFS in node-positive patients only. Because of the short duration of follow-up, the significance of TCD in overall survival was not determined. We conclude that high tumor TCD in node-positive patients is predictive of shorter DFS, and is often associated with HER-2/neu amplification. The possibility exists that high tumor TCD may act in combination with HER-2/neu amplification to promote dissemination of metastases.


Novel Intervertebral Technologies.
Surgical procedures, such as spinal fusion and disk replacement, are commonly used for treatment following failure of conservative treatment in degenerative spine disease. However, there is growing consensus that currently available surgical technologies may have long-term inefficacy for successful management. Intervertebral disk degeneration is the most common manifestation of degenerative spine disease, hence, replacement/repair of this tissue is an important component of surgical treatment. Restoration of spinal alignment and preservation of natural kinematics is also essential to a good outcome. This article reviews novel intervertebral implant technologies that have the potential to significantly impact elective spine surgery for degenerative spine disease.


Epidemiological survey of warts in Chinese military recruits: A cross-sectional and follow-up study.
Warts are very common in military personnel, either at war or during peace times. However, little is known about the prevalence and natural course of warts in military recruits in China. To investigate the prevalence and natural course of warts in Chinese military recruits. In this cross-sectional study, the head, face, neck, hands, and feet of 3093 Chinese military recruits aged 16-25 years in Shanghai were examined for the presence of warts upon enlistment medical examinations. Questionnaires were distributed to collect the general information of the participants before the survey. All the patients were followed up by telephone interview for 11-20 months. The prevalence rate of warts in Chinese military recruits was 2.49%. The diagnosis of most cases was common and plantar warts, which were usually less than 1 cm in diameter and with mild discomfort. Multivariate logistic regression analysis showed that smoking and sharing personal items with others were risk factors. Coming from southern China was a protective factor. Over 2/3 of patients recovered within 1 year and the type, number, and size of warts and treatment choice did not predict resolution.Study limitations and Conclusions This study demonstrated that warts had a relative lower morbidity and a higher spontaneous resolution rate in Chinese military recruits. The telephone interviews following the initial survey and the limitations of a cross-sectional study were the main drawbacks.


Endoribonuclease activities of Trypanosoma brucei mitochondria.
RNA editing in kinetoplastids is a type of post-transcriptional processing that changes mitochondrial mRNA sequences by the addition or deletion of uridines. Multiple enzymatic activities, such as endoribonuclease and RNA ligase, are associated with this process and exist in a multienzyme complex. Endonuclease activities from Trypanosoma brucei mitochondrial extracts were fractionated by sequential ion exchange and gel filtration chromatography. The RNA editing specific endonuclease activity co-fractionated with in vitro editing while another endonuclease activity with a different substrate specificity, and the majority of mtRNase P activity fractionated away from the editing activity. The pH, salt, temperature, and Mg(2+) optima of all three endonucleases were determined. All three activities are sensitive to high temperature and protease digestion. In addition, treatment with micrococcal nuclease resulted in partial disruption of the editing complex and decreased pre-cleaved in vitro insertion editing activity, suggesting that both RNA(s) and protein(s) are necessary in the intact functional complex.


Time-varying effective connectivity of the cortical neuroelectric activity associated with behavioural microsleeps.
An episode of complete failure to respond during an attentive task accompanied by behavioural signs of sleep is called a behavioural microsleep. We proposed a combination of high-resolution EEG and an advanced method for time-varying effective connectivity estimation for reconstructing the temporal evolution of the causal relations between cortical regions when microsleeps occur during a continuous visuomotor task. We found connectivity patterns involving left-right frontal, left-right parietal, and left-frontal/right-parietal connections commencing in the interval [-500; -250] ms prior to the onset of microsleeps and disappearing at the end of the microsleeps. Our results from global graph indices derived from effective connectivity analysis have revealed EEG-based biomarkers of all stages of microsleeps (preceding, onset, pre-recovery, recovery). In particular, this raises the possibility of being able to predict microsleeps in real-world tasks and initiate a 'wake-up' intervention to avert the microsleeps and, hence, prevent injurious and even multi-fatality accidents.


Acute hydrofluoric acid exposure.
Significant local and systemic toxicity may occur from hydrofluoric acid by all routes of exposure. Prompt decontamination by removal from the source and copious irrigation of eyes and skin are essential to reduce morbidity and mortality. Ingestion of small amounts of HF can lead to rapid systemic poisoning and death. Calcium gluconate therapy has become the preferred method of detoxifying the fluoride ion, although its efficacy is based mainly on anecdotal reports and poorly controlled clinical studies. Therefore, more basic research is needed to elucidate the pathophysiology of local toxicity and the best therapeutic modalities to limit injury. All significant exposures should be evaluated by health care personnel familiar with the potential toxicity of this compound.


Greater dietary variety is associated with better biochemical nutritional status in Spanish children: the Four Provinces Study.
Although dietary variety has been associated with a better nutritional profile, its possible role in obesity raises doubts about its overall health benefits. In this study, we examined the association between dietary variety and anthropometric variables, food intake and various food intake biomarkers in Spanish children. This was a cross-sectional study of 1112 children aged 6-7 years from Cadiz, Murcia, Orense and Madrid, who were selected by means of the random cluster-sampling of schools. Information concerning food and nutrient intake was obtained using a food frequency questionnaire, and a dietary variety index (DVI) was calculated on the basis of the number of different foods consumed more than once a month. The anthropometric variables (weight and height), and plasma lipid and vitamin levels were determined using standardised methods. Our results show that the body mass index (BMI) did not vary substantially as a function of DVI: it was 16.9 in the lowest DVI tertile and 17.2 in the highest (p=0.20). Unlike BMI, the DVI positively correlated (p<0.05) with the plasma levels of alpha and beta-carotene, lycopene, retinol, alpha-tocopherol and vitamin E, with energy intake, and with most of the foods, particularly vegetables, fruit and sausages (respective correlation coefficients of 0.43, 0.26 and 0.23). Dietary variety is associated with a better food and nutritional profile in Spanish children. Nevertheless, the presence of a positive association between the DVI and energy intake, and the consumption of sausages and pre-cooked products calls for the recommendation of a varied diet of healthy foods, such as cereals (especially whole grains), fruits and vegetables.


Biological effects of rAAV-caAlk2 coating on structural allograft healing.
Structural bone allografts often fracture due to their lack of osteogenic and remodeling potential. To overcome these limitations, we utilized allografts coated with recombinant adeno-associated virus (rAAV) that mediate in vivo gene transfer. Using beta-galactosidase as a reporter gene, we show that 4-mm murine femoral allografts coated with rAAV-LacZ are capable of transducing adjacent inflammatory cells and osteoblasts in the fracture callus following transplantation. While this LacZ vector had no effect on allograft healing, bone morphogenetic protein signals delivered via rAAV-caAlk2 coating induced endochondral bone formation directly on the cortical surface of the allograft by day 14. By day 28 there was evidence of remodeling of the new woven bone and massive osteoclastic resorption of the cortical surface of the rAAV-caAlk2-coated allografts only. Micro-CT analysis of rAAV-LacZ- vs rAAV-caAlk2-coated allografts after 42 days of healing demonstrated a significant increase in new bone formation (0.67 +/- 0.21 vs 2.49 +/- 0.40 mm(3); P < 0.005). Furthermore, the 3D micro-CT images of femurs grafted with rAAV-Alk2-coated allografts provided the first evidence that complete bridging of bone around a cortical allograft is possible. These results indicate that cell-free, rAAV-coated allografts have the potential to revitalize in vivo following transplantation.


Labeling of unique sequences in double-stranded DNA at sites of vicinal nicks generated by nicking endonucleases.
We describe a new approach for labeling of unique sequences within dsDNA under nondenaturing conditions. The method is based on the site-specific formation of vicinal nicks, which are created by nicking endonucleases (NEases) at specified DNA sites on the same strand within dsDNA. The oligomeric segment flanked by both nicks is then substituted, in a strand displacement reaction, by an oligonucleotide probe that becomes covalently attached to the target site upon subsequent ligation. Monitoring probe hybridization and ligation reactions by electrophoretic mobility retardation assay, we show that selected target sites can be quantitatively labeled with excellent sequence specificity. In these experiments, predominantly probes carrying a target-independent 3' terminal sequence were employed. At target labeling, thus a branched DNA structure known as 3'-flap DNA is obtained. The single-stranded terminus in 3'-flap DNA is then utilized to prime the replication of an externally supplied ssDNA circle in a rolling circle amplification (RCA) reaction. In model experiments with samples comprised of genomic lambda-DNA and human herpes virus 6 type B (HHV-6B) DNA, we have used our labeling method in combination with surface RCA as reporter system to achieve both high sequence specificity of dsDNA targeting and high sensitivity of detection. The method can find applications in sensitive and specific detection of viral duplex DNA.


Low frequency entrainment of oscillatory bursts in hair cells.
Sensitivity of mechanical detection by the inner ear is dependent upon a highly nonlinear response to the applied stimulus. Here we show that a system of differential equations that support a subcritical Hopf bifurcation, with a feedback mechanism that tunes an internal control parameter, captures a wide range of experimental results. The proposed model reproduces the regime in which spontaneous hair bundle oscillations are bistable, with sporadic transitions between the oscillatory and the quiescent state. Furthermore, it is shown, both experimentally and theoretically, that the application of a high-amplitude stimulus to the bistable system can temporarily render it quiescent before recovery of the limit cycle oscillations. Finally, we demonstrate that the application of low-amplitude stimuli can entrain bundle motility either by mode-locking to the spontaneous oscillation or by mode-locking the transition between the quiescent and oscillatory states.


Separation of nucleosomes containing histones H1 and H5.
Hen erythrocyte chromatin was digested with staphylococcal nuclease and fractionated by electrophoresis in polyacrylamide gels. Instead of the three bands described for mouse carcinoma chromatin, four main discrete components (MN1, MN2, MN2E and MN3) were resolved in the mononucleosome fraction of erythrocyte chromatin. MN2 contained all five histones and a DNA fragment of 165--180 base pairs. MN2E comprised four nucleosomal histones plus histone H5 (but not H1) and a DNA fragment of 170--190 base pairs. The relatively nuclease resistant MN3 fraction of erythrocyte nucleosomes contained H1 but no H5 histone. A more accurate analysis of the MN2 fraction in mouse carcinoma nucleosomes revealed some additional microheterogeneity depending on the presence of two different subfractions of H1.


Clinical efficacy of dexamethasone for acute exudative pharyngitis.
The objective of this study was to investigate whether treatment with single-dose dexamethasone can provide relief of symptoms in acute exudative pharyngitis. A prospective, randomized, double-blinded, placebo-controlled clinical trial was undertaken over a 3-month period in a university-based Emergency Department. The study included all consecutive patients between 18 and 65 years of age presenting with acute exudative pharyngitis, sore throat, odynophagia, or a combination, and with more than two Centor criteria. Each patient was empirically treated with azithromycin and paracetamol for 3 days. The effects of placebo and a fixed single dose (8 mg) of intramuscular injection of dexamethasone were compared. The patients were asked to report the exact time to onset of pain relief and time to complete relief of pain. After completion of the treatment, telephone follow-up regarding the relief of pain was conducted. A total of 103 patients were enrolled. Thirty patients with a history of recent antibiotic use, pregnancy, those who were elderly (>65 years of age) and patients who failed to give informed consent were excluded. Forty-two patients were assigned to the placebo group and 31 were assigned to the intramuscular dexamethasone group (8-mg single dose). Time to perceived onset of pain relief was 8.06+/-4.86 h in steroid-treated patients, as opposed to 19.90+/-9.39 h in the control group (p=0.000). The interval required to become pain-free was 28.97+/-12.00 h in the dexamethasone group, vs. 53.74+/-16.23 h in the placebo group (p=0.000). No significant difference was observed in vital signs between the regimens. No side effects and no new complaints attributable to the dexamethasone and azithromycin were observed. Sore throat and odynophagia in patients with acute exudative pharyngitis may respond better to treatment with an 8-mg single dose of intramuscular dexamethasone accompanied by an antibiotic regimen than to antibiotics alone.


Extracellular nucleotides act through P2X7 receptors to activate NF-kappaB in osteoclasts.
Nucleotides, released in response to mechanical and other stimuli, act on P2 receptors in osteoclasts and other cell types. In vitro studies of osteoclasts from rabbits and P2X7 receptor-deficient mice revealed that P2X7 receptors couple to activation of the key transcription factor NF-kappaB. Osteoclasts express functional P2X4 and P2X7 receptors, which are ATP-gated cation channels. Knockout (KO) of the P2X7 receptor has revealed its role in regulating bone formation and resorption, but the underlying signals are not known. The transcription factor NF-kappaB plays a key role in the response of osteoclasts to RANKL and other cytokines. The aim of this study was to examine whether P2X receptors on osteoclasts signal through NF-kappaB. Osteoclasts were isolated from neonatal rabbits or wildtype (WT) and P2X7 receptor KO mice. Immunofluorescence was used to detect the p65 subunit of NF-kappaB, which, on activation, translocates from the cytosol to the nuclei. The concentration of cytosolic free Ca2+ ([Ca2+]i) was monitored in single osteoclasts loaded with fura-2. In control samples, few rabbit osteoclasts demonstrated nuclear localization of NF-kappaB. Benzoyl-benzoyl-ATP (BzATP, a P2X7 agonist, 300 microM) induced nuclear translocation of NF-kappaB after 3 h in approximately 45% of rabbit osteoclasts. In contrast, a low concentration of ATP (10 microM, sufficient to activate P2X4 and P2Y2, but not P2X7 receptors) did not induce nuclear translocation of NF-kappaB. Because BzATP activates multiple P2 receptors, we examined responses of osteoclasts derived from WT and P2X7 receptor KO mice. Treatment with BzATP for 30 minutes increased nuclear localization of NF-kappaB in osteoclasts from WT but not KO mice, showing involvement of P2X7 receptors. Both ATP (10 microM) and BzATP (300 microM) caused transient elevation of [Ca2+]i, indicating that rise of calcium alone is not sufficient to activate NF-kappaB. Pretreatment of rabbit osteoclasts with osteoprotegerin inhibited translocation of NF-kappaB induced by RANKL but not by BzATP, establishing that the effects of BzATP are independent of RANKL signaling. These findings show that P2X7 nucleotide receptors couple to activation of NF-kappaB in osteoclasts. Thus, nucleotides, released at sites of inflammation or in response to mechanical stimuli, may act through NF-kappaB to regulate osteoclast formation and activity.


PPAR-γ agonist pioglitazone prevents apoptosis of endothelial progenitor cells from rat bone marrow.
Selective peroxisome proliferator-activated receptor-γ (PPAR-γ) agonist affects the functions of endothelial progenitor cells (EPCs). This study explores the effect of selective PPAR-γ agonist, pioglitazone, on EPC apoptosis. The cells were cultured and identified via the double staining method in a medium containing different concentrations of pioglitazone. EPC apoptosis was detected by flow cytometry. On Day 7, EPCs engulfed DiL-ac-LDL and FITC-UEA-1, and showed yellow fluorescence in a laser-scanning confocal microscope. EPC apoptosis inhibition was maximal at 50 µmol/L. The ability of pioglitazone to prevent EPC apoptosis may be mediated by the PI3K/Akt signal pathway. The use of thiazolidine two ketone (TZD) to reduce EPC apoptosis may have some potential in treating vascular diseases.


NASA Global Daily Downscaled Projections, CMIP6.
We describe the latest version of the NASA Earth Exchange Global Daily Downscaled Projections (NEX-GDDP-CMIP6). The archive contains downscaled historical and future projections for 1950-2100 based on output from Phase 6 of the Climate Model Intercomparison Project (CMIP6). The downscaled products were produced using a daily variant of the monthly bias correction/spatial disaggregation (BCSD) method and are at 1/4-degree horizontal resolution. Currently, eight variables from five CMIP6 experiments (historical, SSP126, SSP245, SSP370, and SSP585) are provided as procurable from thirty-five global climate models.


Influence of adjuvant hyperbaric oxygen therapy on short-term complications during surgical reconstruction of upper and lower extremity war injuries: retrospective cohort study.
To determine the effects of hyperbaric oxygen (HBO) therapy on short-term complications of complex war wounds to the upper and lower extremities in patients who were and those who were not treated according to North Atlantic Treaty Organization (NATO) emergency war surgery recommendations. We retrospectively analyzed data of 388 male patients undergoing reconstructive surgery for Gustilo type III A, B, and C war wounds to the extremities at the Department of Reconstructive Surgery, Split University Hospital Center, between 1991 and 1995. The occurrence of main wound complications (deep infection, osteomyelitis, skin grafts lyses, and flap necrosis) during hospitalization and time from wounding to granulation formation were analyzed with respect to the use of HBO therapy as a risk factor. Odds ratio (OR) with 95% confidence intervals (CI) was calculated for the occurrence of wound complications with respect to HBO therapy and adjusted for NATO surgical strategy by logistic regression. Of 388 patients, 310 (80%) were initially treated according to the NATO surgical strategy and 99 (25%) received HBO therapy. Deep soft-tissue infection developed in 196 (68%) patients who did not receive HBO therapy and in 35 (35%) who received it (P<0.001, xi(2) test). Osteomyelitis developed in 214 (74%) patients who did not receive HBO therapy and in 62 (63%) who received it (P=0.030). Skin graft lysis occurred in 151 (52%) patients who did not receive HBO therapy and in 23 (23%) who received it (P<0.001). Flap necrosis occurred in 147 (51%) patients who did not receive HBO therapy and in 15 (15%) who received it (P<0.001). Median time to granulation formation was 9 (5-57) days in patients who received HBO therapy, and 12 (1-12) days in those who did not (P<0.001, Mann-Whitney test). These results were consistent over the groups of patients stratified according to the wound severity and remained unaltered after the adjustment for NATO surgical strategy. The effect of HBO therapy was greater in non-NATO than in NATO treated patients in case of deep soft-tissue infection (OR, 10.7 vs OR, 3.8; P=0.031 for interaction). HBO therapy reduced the frequency of wound complications in patients with Gustilo type III wounds and shortened the time to granulation formation. HBO therapy was more effective in non-NATO than in NATO treated patients for the prevention of deep soft-tissue infection but not flap necrosis.


[How large is the loss of effectiveness of a treatment procedure between "theory" and "practice"? Evaluating health economics basic data within the scope of a trial model of ambulatory synchronous balenophototherapy of atopic eczema].
Before introducing new treatment modalities, efficacy has to be proven in clinical trials. Furthermore, it is of special interest to what extent results of clinical trials can be transferred to daily routine. In the framework of a trial on out-patient synchronous balneophototherapy, a possible loss in treatment efficacy from "clinical trial" to "daily routine" was examined. Therefore all patients without protocol deviations (representing treatment success available in a "clinical trial") and all patients with at least one treatment session regardless of protocol deviations (model for "daily routine") were analyzed. 257 "daily routine" and 1281 "clinical trial" patients with atopic dermatitis were analyzed. Relative SCORAD-improvements provided a clear loss in treatment efficacy in the "daily routine" group of 14.2%: 55% of patients received less than the planned 3 treatments per week and 48% left the study early because of noncompliance, good improvement, lack of time or lack of efficacy. For the first time a loss of efficacy of a treatment modality between "theory" and "practice" can be demonstrated in a representative number of patients suffering from atopic dermatitis. For the optimal use of rare financial resources, the comparison of different treatment modalities under "daily routine" conditions is also necessary in future.


Bcl-2 Interacts with Beclin 1 and Regulates Autophagy in 7, 12-Dimethylbenz[a]anthracene-Induced Hamster Buccal-Pouch Squamous-Cell Tumorigenesis.
Autophagy is a programmed cell death procedure, which has essential functions in tumorigenesis. However, its temporal expression and function under different status are yet to be determined. This study aims to investigate the temporal expression of autophagy and its possible function in 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal-pouch cancer model (HBPCM). A total of 50 hamster buccal-pouch tumorigenesis models were established by painting DMBA for 4, 8, 10 and 13 weeks. The expression and subcellular localization of LC3, Beclin 1 and Bcl-2 in buccal lesions were evaluated by immunohistochemical staining and Western blotting. DNA damage was observed by immunohistochemical staining of 8-oHdG. The relationship between Beclin 1 and Bcl-2 was analyzed by immunofluorescence colocalization. The expression levels of LC3 and Beclin 1 associated with autophagy in the experimental buccal pouch of HBPCM were significantly upregulated after 4 weeks (P<0.05), but gradually downregulated after 13 weeks of HBPCM induction. By contrast, the expression level of Bcl-2 was significantly upregulated after 13 weeks. The co-localized regions of Bcl-2 and Beclin 1 peaked after 4 weeks and then decreased gradually. The DNA damage in epithelial cells increased slightly after 4 weeks, and then rapidly decreased over the next 2 months. Autophagy is motivated by a tumor suppressor that diminishes carcinogen-induced DNA damage. However, autophagy is gradually suppressed, which may be attributed to the interaction between Bcl-2 and Beclin 1. This result indicates that the promotion of autophagy may suppress malignant transformation and provide new insights on future potential treatments of HBPCM.


Acute pancreatitis due to severe hypertriglyceridemia in the COVID-19 era: The role of therapeutic plasma exchange.
The psychosocial consequences of the COVID-19 pandemic caused multifaceted challenges in clinical and therapeutic practices. This was the case at the Therapeutic Apheresis Unit of the Padua University Hospital too. Several published reports describe the increase in alcohol and food addiction diseases. In this context, during the last months, the Padua Therapeutic Apheresis Unit treated many more patients with acute pancreatitis due to severe hypertriglyceridemia with therapeutic plasma exchange than in the previous ten years. Furthermore, retrospective cohort studies have been recently published describing the onset of acute pancreatitis during the COVID-19 infection even if, to date, there is still insufficient evidence to estabilish a direct causality. Anyway, the COVID-19 pandemic translated into changes of the overall disease prevalence scenario and therefore the Padua Therapeutic Apheresis Unit will need to reorganise its Therapeutic Apheresis activity.


Effects of heliotropic movements of flowers of Dryas octopetala L. on gynoecium temperature and seed development.
Dryas octopetala has heliotropic flowers, whose petals reflect the light on the pistils, thus warming them up. The effects of this reflection on seed formation and development were studied by measuring the pistil temperature of intact flowers, of flowers with petals removed, of shaded flowers, and of flowers constrained in such a way as always to be directed towards the zenith. In August, the seeds of these flowers were collected, counted and weighed. The temperature differences between the differently treated flowers were greates about noon, the flowers ranking as follows in ascending order of temperature: shaded flowers, flowers without petals, constrained flowers and intact flowers. The mean temperature differences between the gynoecia and the air, in degrees Celsius, were: 1.1, 1.8, 2.5 and 3.2, respectively. The same ranking was obtained for the different treatments, when arranged in order of ascending weight per seed, except that the positions of the shaded flowers and the flowers without petals were interchanged. The mean values of the weight per seed in milligrams were: 0.42, 0.48, 0.53 and 0.61 for the groups flowers without petals, shaded flowers, constrained flowers and intact flowers, respectively.


Measuring proteins on microarrays.
A prerequisite of proteomics is the ability to quantify many selected proteins simultaneously. Immunoassays on microarrays are an attractive solution, as equipment and antibodies are available and assays are simple, scalable and reproducible. Recently, considerable progress has been made in this area as evidenced by increased sensitivity and coverage (degree of multiplexing). Routine use of antibody microarrays in research and diagnostic settings will require increased availability of binding reagents, novel signal amplification procedures, inexpensive and robust platforms for microarray production and detection, and turn-key systems for running high-throughput assays.


Transarterial chemoembolization followed by moderately hypofractionated radiotherapy in hepatocellular carcinoma: Long-term results of RTF3 regimen.
In early-stage hepatocellular carcinoma (HCC) patients merely fit for surgery, transarterial chemoembolization (TACE) achieve low long-term disease control. We evaluated the efficacy and safety of its combination with moderately hypofractionated radiotherapy (hRT) using RTF3 regimen. Between 2006 and 2016, 61 consecutive patients treated in our single expert center for a Barcelona Clinic Liver Cancer (BCLC) A HCC by TACE followed by hRT 3Gy/fraction were retrospectively included. Sixty of the 61 included presented Child-Pugh A cirrhosis (A5, n=41, 67.2%; A6: n=19, 31.1%). Fourteen patients (22.9%) were already treated for a HCC, mainly by radiofrequency (n=12). All patient received a TACE followed by 3Gy per fraction hRT. Mean radiation dose was 54Gy (range: 48-60). After a median follow-up of 118 months, median time-to-progression, progression-free survival (PFS) and overall survival (OS) was 21.3, 18.1, and 31.5 months, respectively. In univariate analysis, PFS was related to dose > 54Gy (HR: 2, P=0.036), and OS was correlated to Child-Pugh A6 or B7 (HR: 1.93, P=0.03) and overall hRT time (HR: 1.06, P=0.015). At progression, orthotopic liver transplantation was performed in 8 patients (13.1%). Severe symptomatic adverse events occurred in 12 patients (19.7%), mainly ascites (n=7). In BCLC-A Child-Pugh A HCC patients ineligible to surgery or thermoablation, TACE-hRT is a safe and effective treatment. Prospective studies are needed to compare this association with radioembolization, TACE-stereotactic radiotherapy, and systemic treatments combinations.


Tomotherapy for cranio-spinal irradiation.
Tomotherapy is a method of delivering rotational IMRT offering various advantages, notably for complex and large targets such as the cranio-spinal axis. This systematic literature review reports on main clinical outcomes and toxicities in patients with various cancer types that received whole craniospinal axis irradiation (CSI) using Tomotherapy and offers a comprehensive comparison between Tomotherapy and other radiotherapy delivery techniques. Databases including PubMed, PubMed Central, Embase, and Cochrane were searched using the keywords "tomotherapy" AND "craniospinal". Fifty-six papers were included in the review. Patient population was adult in 9 papers, paediatric in 26 papers and mixed in 14 papers. Patients treated with helical Tomotherapy had similar disease-specific clinical outcomes and toxicities as patients treated using other techniques. Compared to any other technique, Tomotherapy provides better target coverage, homogeneity, and conformity in 23, 34 and 22 reports. Tomotherapy showed better organ-at-risk sparing for the thyroid, parotids, cochlea, eyes, heart and esophagus. Beam-On-Time (BOT) was reported to be longer for Tomotherapy in most studies (Median BOT: HT = 11 min, VMAT = 5.49 min, 3DCRT = 1.46 min). In conclusion, Tomotherapy offers good cranio-spinal axis coverage with improved homogeneity and conformity compared to other techniques, but with a considerably longer treatment time. Clinical outcome and toxicities suggest using Tomotherapy for CSI is efficient and safe.


Leukoderma punctata.
Thirteen patients with vitiligo (1 segmental, 4 focal, 8 generalized) aged 7 to 38, most of them female children, developed numerous punctate hypopigmented and achromic spots. The spots measured 0.5 to 1.5 mm and were located primarily on the sun-exposed areas of the extremities; they appeared following treatment with PUVASOL. Two of these patients experienced a reduction of this leukodermic defect, whereas the remaining patients showed a stable clinical course. Dopa and Fontana stains disclosed, in most cases, decreased but not absent functional melanocytes and a marked reduction of melanin. Ultrastructural studies demonstrated slight to severe damage of keratinocytes and melanocytes similar to that previously reported in vitiligo patients. The phototoxic effect of PUVASOL therapy is suggested as a possible etiologic factor in these patients. A probable relationship among idiopathic guttate hypomelanosis, leukoderma punctata, and vitiligo is discussed.


Use of Histopaque for isolating mononuclear cells from rabbit blood.
This paper describes the use of Histopaque (Hp) density gradient medium and its advantage for separating mononuclear cells (mnc) from rabbit blood. Hp solutions of density (d) = 1.083 g/ml, 1.119 g/ml and 1:1 mixture of these solutions (final d = 1.103 g/ml) were compared to Ficoll-Paque (Fp, d = 1.077 g/ml) and Lymphopaque (d = 1.086 g/ml). The average leukocyte recovery was: Fp = 26% and Lp = 17%, while that with Hp was: Hp d = 1.083: 41%; Hp d = 1.119: 43%; Hp d = 1.103: 57%. Optimum mnc recovery (76%) was obtained with Hp d = 1.103. Average mnc purity for the various media was: Fp = 88% mnc; Lp = 91%mnc; Hp d = 1.083: 94% mnc; Hp d = 1.119: 93%; Hp d = 1.103: 94% mnc. Contamination was mainly from basophils. Viability was greater than 95% in all cases. Thus, Hp density gradient media provide increased recoveries of mnc from rabbit blood compared to Fp and Lp, while purity is not affected. Rabbit mnc appear to be denser than human mnc, which are recovered in greater numbers using Fp.


[Clinical management of patient with benign prostatic hyperplasia in Spain].
To identify clinical management of benign prostatic hyperplasia (BPH) in Spain and its associated health care resources. A qualitative cross-sectional study was conducted through telephone interviews to general practitioners (GP) and urologists. Information about diagnosis, pharmacologic treatment and follow-up was collected. Results were clustered according to the key variables considered as drivers of clinical practice patterns: BPH diagnosis, severity classification, treatment initiation and follow up of patients. 153 GP and 154 urologists participated in the study. 7 different clinical patterns were identified in primary care (PC). Resource use during diagnosis is relatively homogeneous, reporting a range of 2.0 to 2.6 visits employed and being the most frequent test performed PSA and urine test. Follow-up is heterogeneous; frequency of follow-up visits oscillates from 3.2 to 7.0 visits/patient/year and type of tests performed is different among patterns and within the same pattern. In Urology, 3 clinical patterns were identified. Resource use is homogeneous in the diagnosis and in the follow-up; urologists employed 2 visits in diagnosis and a range of 2.1 to 3.2 visits/patient/year in the follow-up. The most frequent tests both in diagnosis and follow-up are PSA and digital test. BPH management shows variability in PC, identifying 7 different clinical practice patterns with different resource use during the follow-up among patterns and within the same pattern. The implementation of clinical guidelines could be justified to reduce heterogeneity.


Characterization of a new isolate of poliovirus defective interfering particles.
An independent isolate of poliovirus defective interfering particles has been analyzed. These particles, designated DI(A), are apparently analogous to the DI particles described by Baltimore and co-investigators. Electron microscopic heteroduplex analysis reveals that the DI(A) isolate is a mixture of deletion mutants which changes with passage level. The DI(A) population consists of at least five distinct deletion mutants, including one double deletion. Electron microscopic mapping of the deleted regions indicates that most, if not all, of the viral capsid region can be deleted. Despite this heterogeneity, the mutant genomes are quite similar in physical size. We propose a model which suggests that the observed properties of poliovirus DI genomes reflect selective pressures extant during the amplification of the mutant genome. According to this model, only those deleted genomes which retain a minimal size and the capacity to synthesize a functional viral polymerase will replicate successfully in a mixed infection. Furthermore, this model proposes a mechanism for the enrichment of poliovirus DI genomes and an explanation for the low level of complementation observed in mixed infections of picornaviruses.


[Structuro-metabolic aspects of the effect of the adrenal cortex on the pancreas].
In experiments on male rats it was shown that hydrocortisone injection in appropriate doses renders a stimulating effect upon the acinar cell function, whereas large doses of the drug reduce the exocrine tissue secretory function, affecting its protein-synthetizing capacity, first of all. Corticosteroid deficiency is accompanied by morphofunctional atrophy of the exocrine region of the organ. Hydrocortisone and DOCA substituting therapy demonstrates that these changes are caused by glucocorticoid deficiency.


Native T1 mapping in differentiation of normal myocardium from diffuse disease in hypertrophic and dilated cardiomyopathy.
The aim of this study was to examine the value of native and post-contrast T1 relaxation in the differentiation between healthy and diffusely diseased myocardium in 2 model conditions, hypertrophic cardiomyopathy and nonischemic dilated cardiomyopathy. T1 mapping has been proposed as potentially valuable in the quantitative assessment of diffuse myocardial fibrosis, but no studies to date have systematically evaluated its role in the differentiation of healthy myocardium from diffuse disease in a clinical setting. Consecutive subjects undergoing routine clinical cardiac magnetic resonance at King's College London were invited to participate in this study. Groups were based on cardiac magnetic resonance findings and consisted of subjects with known hypertrophic cardiomyopathy (n = 25) and nonischemic dilated cardiomyopathy (n = 27). Thirty normotensive subjects with low pre-test likelihood of cardiomyopathy, not taking any regular medications and with normal cardiac magnetic resonance findings including normal left ventricular mass indexes, served as controls. Single equatorial short-axis slice T1 mapping was performed using a 3-T scanner before and at 10, 20, and 30 minutes after the administration of 0.2 mmol/kg of gadobutrol. T1 values were quantified within the septal myocardium (T1 native), and extracellular volume fractions (ECV) were calculated. T1 native was significantly longer in patients with cardiomyopathy compared with control subjects (p < 0.01). Conversely, post-contrast T1 values were significantly shorter in patients with cardiomyopathy at all time points (p < 0.01). ECV was significantly higher in patients with cardiomyopathy compared with controls at all time points (p < 0.01). Multivariate binary logistic regression revealed that T1 native could differentiate between healthy and diseased myocardium with sensitivity of 100%, specificity of 96%, and diagnostic accuracy of 98% (area under the curve 0.99; 95% confidence interval: 0.96 to 1.00; p < 0.001), whereas post-contrast T1 values and ECV showed lower discriminatory performance. This study demonstrates that native and post-contrast T1 values provide indexes with high diagnostic accuracy for the discrimination of normal and diffusely diseased myocardium.


Acute reduction in functional infarct expansion with late coronary reperfusion: assessment with quantitative two-dimensional echocardiography.
Reperfusion performed too late to salvage myocardium decreases chronic infarct expansion in experimental animals. However, the acute effects of delayed reperfusion are not known. Twenty-two dogs underwent 3 (n = 8), 4 (n = 8) or 6 h (n = 6) of circumflex artery occlusion followed by 3 h of reperfusion. Effects of reperfusion on diastolic expansion were assessed in two ways: 1) change in mean radius of curvature of the infarct segment, and 2) change in the ratio of the length of the diameter from the center of the infarct zone to the opposite wall (septal-lateral diameter) to the length of the diameter perpendicular to this (anteroposterior diameter). Effects on systolic expansion were examined with quantitative two-dimensional echocardiographic systolic thickening analysis. Delayed reperfusion produced an immediate decrease in diastolic infarct expansion. The ratio of septal-lateral/anteroposterior diameters, which had increased with occlusion from a preocclusion baseline of 0.98 +/- 0.06 to 1.13 +/- 0.08 (p less than 0.001), decreased with reperfusion to 1.02 +/- 0.07 at 15 min and 1.03 +/- 0.08 at 3 h of reperfusion (p = 0.001). This was due solely to a decrease in the septal-lateral diameter. The radius of curvature of the infarcted segment increased from 2.1 +/- 0.5 cm before reperfusion to 2.74 +/- 0.8 cm at 15 min and 2.6 +/- 0.85 cm at 3 h of reperfusion (p = 0.009). This occurred despite a significant (13.6%) decline in end-diastolic cavity area and is compatible with flattening of the reperfused infarct region. Systolic infarct expansion also improved slightly.(ABSTRACT TRUNCATED AT 250 WORDS)


Two distinct membrane-bound phosphatidylinositol-4-phosphate phosphatases in bovine brain.
Solubilized phosphatidylinositol-4-phosphate 4-phosphatase from bovine brain resolved into two peaks of activity by ion exchange chromatography. Both exhibited substantial detergent binding characteristic of integral membrane proteins, and both appear specific for phosphatidylinositol-4-phosphate, but their pH optima differ: the earlier eluting fraction (peak 1) is optimally active between pH 5.5 and 6, whereas the later eluting fraction (peak 2) is most active around pH 8.5. Detergent inhibition studies suggest that peak 2, but not peak 1, interacts with phosphatidylinositol-4-phosphate in the context of a single mixed micelle. Further characterization of these activities should help shed light on the biological function of polyphosphoinositide phosphatases.


Effect of Cage-Wash Temperature on the Removal of Infectious Agents from Caging and the Detection of Infectious Agents on the Filters of Animal Bedding-Disposal Cabinets by PCR Analysis.
Efficient, effective cage decontamination and the detection of infection are important to sustainable biosecurity within animal facilities. This study compared the efficacy of cage washing at 110 and 180 °F on preventing pathogen transmission. Soiled cages from mice infected with mouse parvovirus (MPV) and mouse hepatitis virus (MHV) were washed at 110 or 180 °F or were not washed. Sentinels from washed cages did not seroconvert to either virus, whereas sentinels in unwashed cages seroconverted to both agents. Soiled cages from mice harboring MPV, Helicobacter spp., Mycoplasma pulmonis, Syphacia obvelata, and Myocoptes musculinus were washed at 110 or 180 °F or were not washed. Sentinels from washed cages remained pathogen-free, whereas most sentinels in unwashed cages became infected with MPV and S. obvelata. Therefore washing at 110 or 180 °F is sufficient to decontaminate caging and prevent pathogen transmission. We then assessed whether PCR analysis of debris from the bedding disposal cabinet detected pathogens at the facility level. Samples were collected from the prefilter before and after the disposal of bedding from cages housing mice infected with both MPV and MHV. All samples collected before bedding disposal were negative for parvovirus and MHV, and all samples collected afterward were positive for these agents. Furthermore, all samples obtained from the prefilter before the disposal of bedding from multiply infected mice were pathogen-negative, and all those collected afterward were positive for parvovirus, M. pulmonis, S. obvelata, and Myocoptes musculinus. Therefore the debris on the prefilter of bedding-disposal cabinets is useful for pathogen screening.


The effects of psychomotor stimulants on stereotypy and locomotor activity in socially-deprived and control rats.
Using measures of locomotor activity and stereotypy, dose-response curves to several psychomotor stimulant drugs were obtained on rats reared in deprived or normal environments. At both 0.5 and 1.5 mg/kg d-amphetamine, the deprived rats exhibited more intense stereotyped behavior than the control rats. At 5.0 mg/kg, both groups showed maximum response. However, there was no significant difference between the two groups in locomotor response. A similar pattern of results was found for pipradrol, cocaine, and apomorphine. The findings show that different social and environmental experience can modify the response to dopaminergic stimulating agents. The results also suggest that stereotyped behavior should not be considered on the same continuum as locomotor activity. These two behaviors may be mediated by different mechanisms.


An Hspa8 variant is a shocking modifier of spinal muscular atrophy in mice.
In this issue of Neuron, Kim et al.1 show that an Hspa8 variant modifies disease phenotypes in a mouse model of spinal muscular atrophy. Hspa8 facilitates the correct folding of proteins, enhances SNARE assembly, and influences SMN2 splicing.


Identification of and changes in the expression of troponin T isoforms in developing avian and mammalian heart.
Immunochemical studies have shown the presence of a single isoform of troponin T, the adult form, in all chambers of adult chicken and rat hearts. An additional isoform, the embryonic form, was detected during early development of the cardiac muscle. As the amount of adult isoform increased, the embryonic form decreased and was finally suppressed during early post-natal or post-hatch period. The replacement of the embryonic isoform by the adult form appeared slower in the atrium than in the ventricle. While the adult isoform of troponin T was not detected until late in gestation in the rat heart, this isoform of troponin T was present in the chicken heart even at 4 days in ovo.


Fear extinction induced by activation of PKA ameliorates anxiety-like behavior in PTSD mice.
Prolonged exposure (PE) therapy aiming to promote fear extinction is a useful treatment for post-traumatic stress disorder (PTSD). However, the mechanisms underlying fear extinction and effective methods used to promote fear extinction in PTSD are still lacking. In this study, we displayed dysfunctions of cyclic adenosine 3,5-monophosphate (cAMP)-protein kinase A (PKA), protein kinase B (Akt)/mammalian target of rapamycin (mTOR) and calcium signaling in peripheral serum of PTSD patients using bioinformatics analysis. Later, we confirmed the dysfunctions of cAMP-PKA, AKT/mTOR and calcium signaling in the hippocampus of PTSD mice. Moreover, the reduction of calpain1 in the hippocampus enhanced fear memory acquisition. Single activation of PKA by systemic application of rolipram (ROL) or meglumine cyclic adenylate (M-cAMP) before re-exposure promoted fear extinction and improved anxiety-like behavior in PTSD mice. Moreover, systemic application of ROL before re-exposure improved hippocampal brain-derived neurotrophic factor (BDNF)/tyrosine kinase receptor B (TrkB) signaling and calpain1/AKT/mTOR signaling. Interestingly, the effects of activation of PKA could be partially blocked by TrkB antagonist, ANA-12 and mTOR inhibitor, RAPA. Finally, intranasal administration of ROL could also adjust the abnormality of fear memory and improve anxiety-like behaviors in PTSD mice. Collectively, activation of PKA could promote fear extinction, which correlated with the reduction of anxiety-like behavior. The mechanisms were related to the BDNF/TrkB and calpain1/AKT/mTOR signaling pathways. PKA activation might be a useful complementary therapy for PE in the symptom elimination of PTSD.


Geology and fauna of a new Pliocene locality in north-western Kenya.
Lothagam Hill reveals a succession of fossil bearing sediments that extend knowledge of the East African Pliocene back from the base of the Omo succession, to about 5.5 million years.


Long-range structural preformation in yes-associated protein precedes encounter complex formation with TEAD.
Yes-associated protein (YAP) is a partly intrinsically disordered protein (IDP) that plays a major role as the downstream element of the Hippo pathway. Although the structures of the complex between TEA domain transcription factors (TEADs) and the TEAD-binding domain of YAP are already well characterized, its apo state and the binding mechanism with TEADs are still not clearly defined. Here we characterize via a combination of different NMR approaches with site-directed mutagenesis and affinity measurements the intrinsically disordered solution state of apo YAP. Our results provide evidence that the apo state of YAP adopts several compact conformations that may facilitate the formation of the YAP:TEAD complex. The interplay between local secondary structure element preformation and long-range co-stabilization of these structured elements precedes the encounter complex formation with TEAD and we, therefore, propose that TEAD binding proceeds largely via conformational selection of the preformed compact substates displaying at least nanosecond lifetimes.


Bipolaris hawaiiensis-caused phaeohyphomycotic orbitopathy. A devastating fungal sinusitis in an apparently immunocompetent host.
The authors present a case report of a devastating bilateral optic neuropathy and orbitopathy initiated by a contiguous fungal sinusitis, in an apparently immunocompetent young man. The causative organism, Bipolaris hawaiiensis, and other species classified in the genus Bipolaris, are being recognized with increased frequency as causes of several ophthalmic and systemic disorders in both immunocompromised and normal hosts. A literature review demonstrates the variety of clinical presentations with emphasis on those cases which may present to the ophthalmologist.