Citrate Synthase GltA Modulates the 2,4-Diacetylphloroglucinol Biosynthesis of Pseudomonas fluorescens 2P24 and is Essential for the Biocontrol Capacity.
Carbon metabolism is critical for microbial physiology and remarkably affects the outcome of secondary metabolite production. The production of 2,4-diacetylphloroglucinol (2,4-DAPG), a bacterial secondary metabolite with a broad spectrum of antibiotic activity, is a major mechanism used by the soil bacterium Pseudomonas fluorescens 2P24 to inhibit the growth of plant pathogens and control disease occurrence. Strain 2P24 has evolved a complex signaling cascade to regulate the production of 2,4-DAPG. However, the role of the central carbon metabolism in modulating 2,4-DAPG production has not been fully determined. In this study, we report that the gltA gene, which encodes citrate synthase, affects the expression of the 2,4-DAPG biosynthesis gene and is essential for the biocontrol capacity of strain 2P24. Our data showed that the mutation of gltA remarkably decreased the biosynthesis of 2,4-DAPG. Consistent with this result, the addition of citrate in strain 2P24 resulted in increased 2,4-DAPG production and decreased levels of RsmA and RsmE. In comparison with the wild-type strain, the gltA mutant was severely impaired in terms of biocontrol activity against the bacterial wilt disease of tomato plants caused by Ralstonia solanacearum. Moreover, the gltA mutant exhibited increased antioxidant activity, and the expression of oxidative, stress-associated genes, including ahpB, katB, and oxyR, was significantly upregulated in the gltA mutant compared to the wild-type strain. Overall, our data indicate that the citrate synthase GltA plays an important role in the production of 2,4-DAPG and oxidative stress and is required for biocontrol capacity.


Effects of acute uninephrectomy and age on renal blood flow autoregulation in the rat.
We previously showed that renal autoregulatory behavior is modified by both normal and compensatory renal growth. To determine whether the hemodynamic changes precede an increase in renal mass, the relationship of renal blood flow (RBF) to renal perfusion pressure (RPP) was examined in the remaining kidney of anesthetized young and adult rats 30 min following contralateral nephrectomy. In young control animals 32-41 days old, an autoregulation factor (AF) of 0.37 +/- 0.10 (SE) over RPP 70-100 mmHg indicates efficient autoregulation and did not change during the experiment. Acute uninephrectomy in young hydropenic rats resulted in a 16% increase in RBF at normal RPP, and AF increased to 1.22 +/- 0.34 for RPP 70-100 mmHg, reflecting marked impairment of autoregulation. In young animals in which saline infusion was not reduced at the time of uninephrectomy, RBF increased 22% at normal RPP, and autoregulation was similarly impaired (AF 0.92 +/- 0.12 for RPP 70-100 mmHg). Acute uninephrectomy in adult rats resulted in a 13% increase in RBF, with better preservation of AF (0.48 +/- 0.09) over the adult autoregulatory range (RPP 100-130 mmHg). These findings indicate that profound hemodynamic changes take place in the remaining kidney within 30 min of acute uninephrectomy and that they are qualitatively similar to those observed after compensatory hypertrophy has developed.


Smartphone-Assisted Medical Care for Vestibular Dysfunction as a Telehealth Strategy for Digital Therapy Beyond COVID-19: Scoping Review.
Dizziness and vertigo can be caused by various factors, such as peripheral vestibular and central disorders. Although consultations with specialists are advisable when necessary, patients with severe vertigo symptoms may have limited mobility, which may interfere with hospital visits. The spread of COVID-19 has further limited the number of hospital visits for patients with dizziness; therefore, a method of medical care that enables more accurate treatment under time and geographical constraints is needed. Telemedicine has become widespread, owing to the popularity of smartphone and tablet devices in recent years, and the use of devices and systems has made it possible to provide efficient medical care. However, no previous scoping review has mapped existing studies on telemedicine for vertigo and dizziness, and no recommendations have been made regarding which devices and systems should be used for specific diseases. The aim of this review was to map and assess previous studies on the use of information communications technology, smartphones, and apps for treating patients with vertigo and discuss the added value of introducing telemedicine to improve the quality of medical care and create an environment that builds security and trust among patients. A scoping review was conducted with the methodological framework of Arksey and O'Malley and in accordance with the of the PRISMA-ScR (Preferred Reporting Items for Systematic Reviews and Meta-Analysis extension for Scoping Reviews) guidelines. The PubMed, MEDLINE, and Cochrane Library databases were searched to retrieve previous reports on smartphone-assisted telemedicine treatment for vertigo published between January 2000 and May 2023. Two authors independently assessed eligibility and extracted data. This review included 20 papers that reported devices or systems for telemedicine for vestibular dysfunction. Among studies that reported the use of a device or app, 2 were related to anamnesis and subjective symptoms, 12 were related to objective examination, 7 were related to remote diagnosis, and 7 were related to treatment and rehabilitation. With the advancement of technology, the use of telemedicine in patients with dizziness may be feasible. In the future, it will be necessary to consider how telemedicine can be used in dizziness treatment and develop an effective treatment system combining in-person medical care and the effective use of devices for the management of severe vertigo and related diseases. The smooth introduction of telemedicine in vertigo treatment is expected to improve the quality of treatment, increase opportunities for patients to receive medical care, and reduce time and travel costs, leading to a sense of security and trust among patients.


Genomic regions required for morphogenesis of the Drosophila embryonic midgut.
The Drosophila midgut is an excellent system for studying the cell migration, cell-cell communication, and morphogenetic events that occur in organ formation. Genes representative of regulatory gene families common to all animals, including homeotic, TGF beta, and Wnt genes, play roles in midgut development. To find additional regulators of midgut morphogenesis, we screened a set of genomic deficiencies for midgut phenotypes. Fifteen genomic intervals necessary for proper midgut morphogenesis were identified, three contain genes already known to act in the midgut. Three other genomic regions are required for formation of the endoderm or visceral mesoderm components of the midgut. Nine regions are required for proper formation of the midgut constrictions. The E75 ecdysone-induced gene, which encodes a nuclear receptor superfamily member, is the relevant gene in one region and is essential for proper formation of midgut constrictions. E75 acts downstream of the previously known constriction regulators or in parallel. Temporal hormonal control may therefore work in conjunction with spatial regulation by the homeotic genes in midgut development. Another genomic region is required to activate transcription of the homeotic genes Antp and Scr specifically in visceral mesoderm. The genomic regions identified by this screen provide a map to novel midgut development regulators.


Modeling regulatory cascades using Artificial Neural Networks: the case of transcriptional regulatory networks shaped during the yeast stress response.
Over the last decade, numerous computational methods have been developed in order to infer and model biological networks. Transcriptional networks in particular have attracted significant attention due to their critical role in cell survival. The majority of network inference methods use genome-wide experimental data to search for modules of genes with coherent expression profiles and common regulators, often ignoring the multi-layer structure of transcriptional cascades. Modeling methodologies on the other hand assume a given network structure and vary significantly in their algorithmic approach, ranging from over-simplified representations (e.g., Boolean networks) to detailed -but computationally expensive-network simulations (e.g., with differential equations). In this work we use Artificial Neural Networks (ANNs) to model transcriptional regulatory cascades that emerge during the stress response in Saccharomyces cerevisiae and extend in three layers. We confine the structure of the ANNs to match the structure of the biological networks as determined by gene expression, DNA-protein interaction and experimental evidence provided in publicly available databases. Trained ANNs are able to predict the expression profile of 11 target genes across multiple experimental conditions with a correlation coefficient >0.7. When time-dependent interactions between upstream transcription factors (TFs) and their indirect targets are also included in the ANNs, accurate predictions are achieved for 30/34 target genes. Moreover, heterodimer formation is taken into account. We show that ANNs can be used to (1) accurately predict the expression of downstream genes in a 3-layer transcriptional cascade based on the expression of their indirect regulators and (2) infer the condition- and time-dependent activity of various TFs as well as during heterodimer formation. We show that a three-layer regulatory cascade whose structure is determined by co-expressed gene modules and their regulators can successfully be modeled using ANNs with a similar configuration.


Differential staining of different types of peripheral nerve and muscle.
We modified Ungewitter's (1951) and Beermann and Cassens' (1976) non-selective silver methods for peripheral nerve axons and their terminals by lengthening the time for the prestaining treatment of sections, adjusting the times of various stages of the staining procedures and selecting reagents with minimal chloride and sulphate impurities. These methods were applied to Bouin's fixed material embedded in wax with serial sections up to 100 micron thick, cut longitudinally. The Ungewitter modification stained sensory axons and their terminals very well but failed to give good impregnations of motor fibres. In addition, it marked intrafusal muscle fibres with a peppery silver deposit while sparing extrafusal fibres. The Beermann and Cassens method yielded excellent details of motor axons and their terminals but results were very poor on sensory fibres. Both sensory and motor methods worked regardless of whether the neural elements were mature, regenerating, or neonatal. Both methods consistently yielded pale backgrounds in these thick sections.


Correction of post-traumatic flexion contracture of the elbow by anterior capsulotomy.
Despite prolonged conservative therapy, including splinting, bracing, and supervised exercise programs, significant flexion contracture of the elbow after local trauma may remain both a functional and a cosmetic impairment. We studied the cases of fifteen patients who underwent a relatively limited surgical procedure consisting of anterior capsulotomy of the elbow without tenotomy of the biceps tendon or myotomy of the brachialis muscle. The mean preoperative flexion deformity of 48 degrees was reduced to a mean postoperative deformity of 19 degrees (a 61 per cent improvement). Two groups of patients were identified. Eleven patients who did not have significant preoperative evidence of intra-articular degenerative (post-traumatic) changes in the joint achieved a 65 per cent improvement in extension, with one patient having continued pain with vigorous activity. Four patients who had significant preoperative post-traumatic arthrosis achieved a 50 per cent improvement in extension; however, three of these four patients continued to have significant pain. Limited anterior capsulotomy is a safe and effective means of decreasing post-traumatic flexion contracture of the elbow in properly selected patients. Better overall results are obtained in elbows without significant post-traumatic arthrosis.


Ethical aspects when treating traumatized refugee children and their families.
Basic ethical principles are worth analysing step by step when dealing with refugee children and their families. Three issues where potential ethical conflicts might arise for healthcare professionals in treating refugees with different cultural background are pointed out--traumatic life events, hierarchy and repatriation. An ethical analysis of the decision to admit a traumatized teenage refugee to a psychiatric ward is discussed with respect for the ethical principles autonomy, non-maleficence, beneficence and justice. There are both gains and losses, which are valued differently depending on the actors involved.


A pedicled split extended vertical deep inferior epigastric (s-vDIEP) flap and an adipo-dermal thigh local flap for the reconstruction of the medial thigh compartment after sarcoma resection: A case report.
Large upper medial thigh defects in prior irradiated tissue require challenging reconstructions. Several techniques have been reported to reconstruct this region and according to the literature, pedicled perforator flaps are the first reconstructive option. The anterolateral thigh flap is considered the gold standard, while surprisingly the pedicle deep inferior epigastric (DIEP) flap in vertical fashion has not been frequently employed, if compared with its muscular counterpart, the pedicle vertical rectus abdomins flap (vRAM). We report a case of a multilayered flaps reconstruction of the left medial thigh after an excision of a sarcoma involving the whole adductors compartment. A 75-year-old male patient underwent a free margins resection of the sarcoma. After the resection, a soft tissue defect of 24 cm × 14 cm × 14 cm spreading from the groin to the medial tuberosity of the tibia, was left. We performed a reconstructive technique based on a pedicled split extended vertical deep inferior epigastric (s-vDIEP) flap and an adipo-dermal thigh local flap in order to fill and cover the thigh defect. The s-vDIEP had 2 islands: a cranial de-epithelialized island to fill the dead space and a caudal for the skin closure. The postoperative follow-up was complicated by seroma formation and it was managed by sclerotherapy and at the 6 months follow-up the patient showed good cosmetic and functional outcomes with no sign of tumor relapse. Our result suggests that the proposed multilayered reconstruction may be employed for the restoration of large and deep upper medial thigh defects.


Propionibacterium species and follicular keratinocyte activation in acneic and normal skin.
The pathogenesis of acne vulgaris is multifactorial with increased sebum production, alteration in the quality of sebum lipids, dysregulation of the hormone microenvironment, follicular hyperkeratinization and Propionibacterium acnes-driven inflammation as major contributory factors. Hyperproliferation of keratinocytes is believed to contribute to hypercornification and eventually leads to comedone development. While the distribution of P. acnes is relatively well documented in acneic and healthy skin, little is known about P. granulosum and P. avidum. To visualize directly the three major Propionibacterium in 117 control and 26 acneic skin samples. In addition, keratinocyte proliferation was evaluated. Propionibacteria were visualized by immunofluorescence microscopy, and keratinocyte proliferation was assessed by Ki67, keratin (K) 16 and p63 immunochemistry. P. acnes was identified in 68 samples (48%), while P. granulosum was identified in 12 (8%) samples; P. avidum was not detected at all. Unexpectedly, acne samples did not show higher keratinocyte proliferation than controls, nor was there any association between bacterial colonization and expression of Ki67/K16/p63. Our findings do not support earlier notions of follicular keratinocyte hyperproliferation as a cause of ductal hypercornification in acneic facial skin. Further studies on the mechanisms underlying hypercornification in acne pathogenesis are needed.


Autophagy in the regulation of pathogen replication and adaptive immunity.
Autophagy is an evolutionarily conserved homeostatic process by which cells deliver cytoplasmic material for degradation into lysosomes. Autophagy may have evolved as a nutrient-providing homeostatic pathway induced upon starvation, but with the acquisition of cargo receptors, autophagy has become an important cellular defence mechanism as well as a generator of antigenic peptides for major histocompatibility complex (MHC) presentation. We propose that autophagy efficiently protects against microbes encountering the cytosolic environment accidentally, for example, upon phagosomal damage, whereas pathogens routinely accessing the host cytosol have evolved to avoid or even benefit from autophagy.


Environmental effects on the autoxidation of retinol.
1. The behaviour of retinol in aqueous colloidal dispersions has been studied because, if membranes are a physiological site of action of vitamin A, the reactions of colloidal retinol may be relevant to the functions of the vitamin in vivo. 2. Dispersions of retinol in NaCl exhibit characteristic spectral changes, and they consume O(2), within minutes of preparation. 3. The maximum rate of O(2) uptake is approximately linearly dependent on the concentration of O(2). 4. At limiting concentrations of O(2), the spectral changes are accelerated by catalase, indicating that H(2)O(2) is one of the reaction products. 5. The autoxidation, which is relatively unaffected by light, has the characteristics of a radical-catalysed reaction. O(2) uptake is preceded by an exceptionally short induction period; the reaction is catalysed by Fe(2+) ions and is inhibited by diphenylpicrylhydrazyl. 6. The maximum rate of autoxidation, which is less in water or sucrose solution than in saline, depends on the degree of aggregation of retinol molecules induced by cations. 7. In the absence of O(2), the cation-induced aggregates exhibit a spectral red-shift, which difference-spectra indicate is caused by formation of a species with lambda(max.) 370-380nm. 8. This species, from which retinol can be quantitatively recovered, is apparently the oxygen-sensitive form of retinol that initiates the rapid autoxidation. 9. The possible biological significance of the production of a highly reactive form of retinol in micellar aggregates is discussed.


Antiglycation Effects of Adlay Seed and Its Active Polyphenol Compounds: An In Vitro Study.
This study aimed to evaluate the antiglycation effects of adlay on protein glycation using in vitro glycation assays. Adlay seed was divided into the following four parts: the hull (AH), testa (AT), bran (AB), and polished adlay (PA). A solvent extraction technique and column chromatography were utilized to investigate the active fractions and components of adlay. Based on a BSA-glucose assay, the ethanolic extracts of AT (ATE) and AB (ABE) revealed a greater capacity to inhibit protein glycation. ATE was further consecutively partitioned into four solvent fractions with n-hexane, ethyl acetate (ATE-Ea), 1-butanol (ATE-BuOH), and water. ATE-BuOH and -Ea show marked inhibition of glucose-mediated glycation. Medium-high polarity subfractions eluted from ATE-BuOH below 50% methanol with Diaion HP-20, ATE-BuOH-c to -f, exhibited superior antiglycation activity, with a maximum inhibitory percentage of 88%. Two phenolic compounds, chlorogenic acid and ferulic acid, identified in ATE-BuOH with HPLC, exhibited potent inhibition of the individual stage of protein glycation and its subsequent crosslinking, as evaluated by the BSA-glucose assay, BS-methylglyoxal (MGO) assay, and G.K. peptide-ribose assay. In conclusion, this study demonstrated the antiglycation properties of ATE in vitro that suggest a beneficial effect in targeting hyperglycemia-mediated protein modification.


Posttranslational modifications define course of prion strain adaptation and disease phenotype.
Posttranslational modifications are a common feature of proteins associated with neurodegenerative diseases including prion protein (PrPC), tau, and α-synuclein. Alternative self-propagating protein states or strains give rise to different disease phenotypes and display strain-specific subsets of posttranslational modifications. The relationships between strain-specific structure, posttranslational modifications, and disease phenotype are poorly understood. We previously reported that among hundreds of PrPC sialoglycoforms expressed by a cell, individual prion strains recruited PrPC molecules selectively, according to the sialylation status of their N-linked glycans. Here we report that transmission of a prion strain to a new host is accompanied by a dramatic shift in the selectivity of recruitment of PrPC sialoglycoforms, giving rise to a self-propagating scrapie isoform (PrPSc) with a unique sialoglycoform signature and disease phenotype. The newly emerged strain has the shortest incubation time to disease and is characterized by colocalization of PrPSc with microglia and a very profound proinflammatory response, features that are linked to a unique sialoglycoform composition of PrPSc. The current work provides experimental support for the hypothesis that strain-specific patterns of PrPSc sialoglycoforms formed as a result of selective recruitment dictate strain-specific disease phenotypes. This work suggests a causative relationship between a strain-specific structure, posttranslational modifications, and disease phenotype.


Cross-cultural adaptation and measurement properties of the Brazilian-Portuguese version of the Cincinnati Knee Rating System.
Purpose: To translate, culturally adapt, evaluate the measurement properties, and propose a new scoring system of the Cincinnati Knee Rating System for the Brazilian population.Materials and Methods: One hundred fifty people with anterior cruciate ligament injury completed Cincinnati Knee Rating System and Lysholm Knee Scoring Scale on three occasions: 1-week prior to surgery, 90-days after and 95-days after reconstruction. The measurement properties of the Brazilian-Portuguese Cincinnati Knee Rating System, internal consistency, construct validity, reproducibility, ceiling and floor effect and responsiveness, were tested.Results: Using the original scoring system (multiple scores), the internal consistency (Cronbach's α) varied between 0.54-0.79 (if item deleted = 0.07-0.73); the construct validity (Pearson's r) varied between 0.19-0.82 (related to Lysholm); the reliability (intraclass correlation coefficient) varied between 0.96-0.99; the standard error of measurement varied between 0.2-1.3 points; the minimum detectable change varied between 0.4-3.5 points; no ceiling or floor effect was detected, and responsiveness (effect size) varied between -0.3-2.7. Using the new proposed scoring system (single score), Cronbach's α was 0.78 (if item deleted = 0.65-0.71); the Pearson's r was 0.79 (related to Lysholm); the intraclass correlation coefficient was 0.99; the standard error of measurement was 0.5 points, the minimum detectable change was 1.3 points; no ceiling or floor effect was detected, and effect size was 1.4.Conclusions: The Brazilian-Portuguese Cincinnati Knee Rating System has adequate property measurement and can be used in a Brazilian population. The new proposed scoring system is appropriate.Implications for rehabilitationThe Brazilian-Portuguese Cincinnati Knee Rating System is a valid and reliable instrument which can identify consistent clinical changes over the time.The Brazilian-Portuguese Cincinnati Knee Rating System can be used to evaluate and to follow-up a Brazilian population with anterior cruciate ligament injuries and postoperative reconstruction.The new proposed score for Cincinnati Knee Rating System is appropriate and did not compromise the quality of the measurement properties.The new scoring system generates a unique score, creating a potentially simpler and faster clinical understanding of the patient's condition.


Acetaminophen-induced proliferation of breast cancer cells involves estrogen receptors.
Previous studies have shown that acetaminophen, a common analgesic/antipyretic, induces proliferation of cultured breast cancer cells containing both estrogen and progesterone receptors (ER+/PR+). The main objective of this study was to evaluate the involvement of ERs in this effect. First, the effects of therapeutic acetaminophen concentrations were compared in breast cancer cells with high ERs and in T47Dco cells with lower ERs, to determine if acetaminophen-induced proliferation depends on ER levels. Second, the effects of two antiestrogens (ICI 182,780 and 4'-hydroxytamoxifen) on acetaminophen-induced proliferation were determined in three human breast cancer cell lines: two ER+/PR+ (MCF7, T47D) and one ER-/PR- (MDA-MB-231). Third, ER binding assays were performed in MCF7 cells to determine if acetaminophen competed with estradiol for binding to ERs. Proliferation endpoints monitored included percent cells in the DNA synthesis phase of the cell cycle, 3H-thymidine incorporation into DNA, and cell number. Acetaminophen did not induce DNA synthesis in T47Dco cells, but did in cells with higher ER levels, suggesting high ER levels are necessary for acetaminophen to induce proliferation. Antiestrogens inhibited acetaminophen-induced proliferation in ER+/PR+ cells while no effects were observed in ER-/PR- cells, further supporting ER involvement. However, acetaminophen did not compete with estradiol for binding to ERs in ER+/PR+ cells. Collectively, these data suggest that acetaminophen induces breast cancer cell proliferation via ERs without binding to ERs like estradiol. The second purpose of this study was to determine if acetaminophen is estrogenic/antiestrogenic in vivo (uterotrophic assays). Acetaminophen has no antiestrogenic/estrogenic activity in mice or rats uteri.


How neurons generate behavior in a hatchling amphibian tadpole: an outline.
Adult nervous systems are so complex that understanding how they produce behavior remains a real challenge. We chose to study hatchling Xenopus tadpoles where behavior is controlled by a few thousand neurons but there is a very limited number of types of neuron. Young tadpoles can flex, swim away, adjust their trajectory, speed-up and slow-down, stop when they contact support and struggle when grasped. They are sensitive to touch, pressure, noxious stimuli, light intensity and water currents. Using whole-cell recording has led to rapid progress in understanding central networks controlling behavior. Our methods are illustrated by an analysis of the flexion reflex to skin touch. We then define the seven types of neuron that allow the tadpole to swim when the skin is touched and use paired recordings to investigate neuron properties, synaptic connections and activity patterns. Proposals on how the swim network operates are evaluated by experiment and network modeling. We then examine GABAergic inhibitory pathways that control swimming but also produce tonic inhibition to reduce responsiveness when the tadpole is at rest. Finally, we analyze the strong alternating struggling movements the tadpole makes when grasped. We show that the mechanisms for rhythm generation here are very different to those during swimming. Although much remains to be explained, study of this simple vertebrate has uncovered basic principles about the function and organization of vertebrate nervous systems.


DTI at long diffusion time improves fiber tracking.
While diffusion-tensor-imaging tractography provides remarkable in vivo anatomical connectivity of the central nervous system, the majority of DTI studies to date are predominantly limited to tracking large white-matter fibers. This study investigated DTI tractography using long diffusion time (t(diff)) to improve tracking of thinner fibers in fixed rhesus monkey brains. Stimulated Echo Acquisition Mode (STEAM) sequence on a 3T Siemens TRIO was modified to include a diffusion module. DTI was acquired using STEAM with t(diff) of 48 and 192 ms with matched signal-to-noise ratios (SNR). Comparisons were also made with the conventional double-spin echo (DSE) at a short t(diff) of 45 ms. Not only did the fractional anisotropy increase significantly with the use of long diffusion time, but directional entropy measures indicated that there was an increased coherence amongst neighboring tensors. Further, the magnitude of the major eigenvector was larger at the t(diff) = 192 ms as compared to the short t(diff). Probabilistic connectivity maps at long t(diff) showed larger areas of connectivity with the use of long diffusion time, which traversed deeper into areas of low anisotropy. With tractography, it was found that the length of the fibers, increased by almost 10% in the callosal fibers that branch into the paracentral gyrus, the precentral gyrus and the post central gyrus. A similar increase of about 20% was observed in the fibers of the internal capsule. These findings offer encouraging data that DTI at long diffusion time could improve tract tracing of small fibers in areas of low fractional anisotropy (FA), such as at the interfaces of white matter and grey matter.


Activation or suppression of the immune response mediators in biliary tract cancer (BTC) patients: a systematic review and meta-analysis.
Background: Infiltration of immune cells and immune microenvironment determine the proliferative activity of the tumor and metastasis. The aim of this study was to analyze the influence of activation or suppression of the immune response mediators on the prognosis of biliary tract cancer (BTC). Methods: We searched Pubmed, Web of Science, Embase and The Cochrane Library for relevant literatures until June 2016. The quality of studies was assessed by QUADAS-2 and NOS tools. Forest and funnel plots and all statistical analyses were generated by using Review Manager 5.3. The bias of included studies was estimated by Egger's test using Meta R package. Results: A total of 2339 patients from 12 studies were finally enrolled in this meta-analysis. Patients with high expression of immune active factors, intraepithelial tumor-infiltrating CD4+ , CD8+, and Foxp3+ T lymphocytes, MHC I, NKG2D, showed a better overall survival (OS) than those with low expression (HR=0.52, 95% CI=0.41-0.67, P<0.00001). On the contrary, the high expression of immune suppressive factors (CD66b+ neutrophils, Neutrophil-lymphocyte ratio, Intratumoral IL-17+ cells and PD-1+/CD8+ TILs) was significantly associated with poor OS (HR=1.79, 95% CI=1.44-2.22, P<0.00001). A further analysis of therapies targeting tumor microenvironment modulation showed that the median progression free survival (PFS) for BTC patients who received adjuvant immunotherapy was longer than those who received surgery or chemotherapy alone, and the estimated pooled mean difference demonstrated a highly significant improvement (MD =2.33; 95% CI: 0.63-4.02, P=0.007). The total effect of PFS and OS was statistically longer in experimental group, compared to patients in control groups, respectively (PFS: RR=1.25; 95% CI: 1.08-1.46, P=0.004; OS: RR=1.16; 95% CI: 1.07-1.27, P=0.0006). In subgroup meta-analysis of studies on 6-, 12- and 18-month PFS and OS, it showed that adjuvant immunotherapy could improve the 6-month PFS (RR=1.23; 95% CI: 1.05-1.44, P=0.009), and 6-month OS (RR=1.17; 95% CI: 1.06-1.30, P=0.002). Conclusions: So given the above issue, our meta-analysis confirmed that the level of immune mediators could be a predicative factor for prognosis of BTC patients, and immunotherapy regimens by modulating the tumor microenvironment was superior for enhancing median PFS, 6-month PFS and OS.


Pharmacogenetic effects of 'candidate gene complexes' on stroke in the GenHAT study.
The aim of this study was to investigate whether there is a genotype-by-treatment interaction in patients experiencing stroke and treated with one of three antihypertensive drugs, that is chlorthalidone, amlodipine, or lisinopril. A population of 436 African Americans and 539 whites who had experienced stroke in the GenHAT study were genotyped for 768 single nucleotide polymorphisms (SNPs) in 280 candidate genes. To detect a genotype-by-treatment interaction, we used the Pearson's χ-test to assess whether the genotype frequencies differed at the single SNP level for the three drug treatment groups. From these single SNP analyses, we derived a summary statistic for the degree of association at the gene and gene complex levels. This was done by grouping SNPs using information on gene locations and defining gene complexes on the basis of protein-protein interactions. To assess the statistical significance of the observed test statistic, we derived an empirical P-value by simulating data under the null hypothesis. We found that, in patients who have experienced stroke, there is a significant genetic difference between hypertension drug treatment groups. In African Americans, SNP rs12143842 showed a significant association (P<0.001) with drug treatment. At the gene level, HNRNPA1P4 and NOS1AP in African Americans and PRICKLE1 and NINJ2 in non-Hispanic whites were significantly associated (P<0.01) with drug treatment, whereas none of the gene complexes tested showed significance. On the basis of the genetic differences between drug treatment groups, we conclude that there may be an interaction between certain genotypes and antihypertensive treatment in stroke patients. This needs to be replicated in other studies.


Physical characteristics of the ECAT EXACT3D positron tomograph.
The 'EXACT3D' positron tomograph, which is now in routine clinical research use, was developed with the aim of achieving unprecedented sensitivity, high spatial and temporal resolution and simplicity of design using proven detector technology. It consists of six rings of standard detector blocks (CTI/Siemens EXACT HR+) with 4.39 mm x 4.05 mm x 30 mm elements, giving an axial field of view (FOV) of 23.4 cm. This extended FOV and the absence of interplane septa and retractable transmission rod sources has allowed greatly simplified gantry and detector cassette design. Operation in exclusive 3D mode requires an alternative to the conventional coincidence method for transmission scanning, and a single photon approach using a hydraulically driven 137Cs point source has been implemented. The tomograph has no other moving parts. A single time frame of data without any compression is very large (> 300 Mbyte) and two approaches are employed to overcome this difficulty: (a) adjacent sinograms can be summed automatically into different combinations and (b) listmode (event-by-event) acquisition has been instituted, which is both storage efficient (particularly for acquisition of sparse data sets) and maximizes temporal resolution. The high-speed I/O and computing hardware can maintain a sustained acquisition rate of about 4 million coincidence events per second. A disadvantage of the large axial FOV in 3D is the increased sensitivity to activity outside the coincidence FOV. However, this can be minimized by additional side shielding. The mean spatial resolution is 4.8 +/- 0.2 mm FWHM (transaxial, 1 cm off-axis) and 5.6 +/- 0.5 mm (axial, on-axis). Its absolute efficiency is 5.8% for a line source in air (just spanning the axial FOV) and 10% for a central point source (with thresholds of 350-650 keV). For a uniform 20 cm diameter cylinder, the efficiency is 69 kcps kBq(-1) ml(-1) (after subtraction of a scatter fraction of 42%). Sensitivity relative to the EXACT HR+ (with four rings of blocks) is 2.5 (3D) and 12 (2D) times respectively. The rate of random events in blood flow studies in the brain and body, using 15O-labelled water, can be controlled by limiting the administered dose and inserting additional side shielding.


Layer-specific somatosensory cortical activation during active tactile discrimination.
Ensemble neuronal activity was recorded in each layer of the whisker area of the primary somatosensory cortex (SI) while rats performed a whisker-dependent tactile discrimination task. Comparison of this activity with SI activity evoked by similar passive whisker stimulation revealed fundamental differences in tactile signal processing during active and passive stimulation. Moreover, significant layer-specific functional differences in SI activity were observed during active discrimination. These differences could not be explained solely by variations in ascending thalamocortical input to SI. Instead, these results suggest that top-down influences during active discrimination may alter the overall functional nature of SI as well as layer-specific mechanisms of tactile processing.


Flare as applied to photographic lenses.
This paper is concerned with flare measurement in photographic lenses by three colors instead of white light. The results have interesting relations to the antireflection coating, the barrel interior finishing, area and condition of the iris diaphragm of the lens, and the lens edge painting. For color photography, it is more realistic to evaluate flare by means of the relative amount present in each of the blue, green, and red color components. In accordance with the data thus measured, various countermeasures can be considered.


[Analysis of current status of eye bank construction in Western China].
To evaluate the current status of eye bank construction in Western China. The numbers of eye banks, equipments of eye banks, methods of donor cornea preservation, fund for managing eye banks, technicians of eye banks, and difficulties in the construction of eye bank were surveyed by questionnaires from ophthalmology associations and eye banks in the following provinces and city: Sichuan province, Yunnan province, Guizhou province, Guangxi Zhuang autonomous region, Tibet autonomous region, Shanxi province, Gansu province, Ningxia Hui autonomous region, Qinghai province, Xinjiang Weiwuer autonomous region, Neimonggu autonomous region, and city of Chongqing before March 2003, and the numbers of eye banks and donor corneas obtained from January 1999 to March 2003 were also included in the study. The current situation of eye bank construction in Western China showed: (1) There were total 8 eye banks in 6 provinces among the 12 provinces and city of Western China. The mean coverage rate of eye bank in Western China was 50%. There were two eye banks in Chongqing and Shanxi separately, one eye bank in Sichuan, Yunnan, Gansu and Xinjiang respectively. (2) The fundamental equipment of each eye bank included super-clean worktable, slit-lamp microscope, refrigerator, and liquid nitrogen jar. There were 5 specular microscopes and 6 computers in the 8 eye banks of Western China. The moist chamber and medium-term corneal preserving medium were used as short term and medium-term storage methods to keep the endothelial cell survival of donor cornea. As a long term preservation of corneal donor, glycerine dehydration and deep low temperature were still used regularly. (3) financial support of eye bank was from hospital, government assigned fund, international Orbis organization and etc. (4) There were only one or two technicians in majority of the eye banks investigated in the study. (5) From January 1999 to March 2003, community education regarding to donation of eye was carried out forty-two times. During the same period, the donated cornea was added up to 156, and 126 of them were used in clinical transplantation. The difficulties in the construction of eye banks in Western China included: the amount of donated cornea was not enough; the technical exchange and cooperation between the eye banks were little; the patients could not afford the cost of transplantation surgery. The current situation of eye bank construction in Western China was behind that of rest areas in China, and the development of eye banks in Western China was not equal.


Microenvironmental pH-modified solid dispersions to enhance the dissolution and bioavailability of poorly water-soluble weakly basic GT0918, a developing anti-prostate cancer drug: preparation, characterization and evaluation in vivo.
The aim of the present work was to design a pH-modified solid dispersion (pH(M)-SD) that can improve the dissolution and bioavailability of poorly water-soluble weakly basic GT0918, a developing anti-prostate cancer drug. To select the appropriate acidifiers, a solubility test was carried out first. Solid dispersions (SDs) containing GT0918 and polyvinylpyrrolidone (PVP) were prepared using a solvent evaporation method and were characterized using dissolution studies in different media. The solid states of the SDs were investigated using scanning electron microscopy (SEM), powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC) and Fourier transformed infrared spectroscopy (FTIR). The in vivo pharmacokinetics of the pH(M)-SDs tablets were also studied in beagle dogs compared to the conventional tablets. The optimized pH(M)-SD (GT0918/PVP/citric acid, 1:2:2 weight ratio) exhibited a significant improvement in the dissolution behavior compared to both the physical mixture and the binary SDs. Solid-state characterization revealed that the amorphous formation of GT0918 in the SDs and the strong H-bonding were only found in the pH(M)-SDs containing citric acid. Furthermore, the GT0918-loaded pH(M)-SD tablets showed a higher AUC and a lower tmax compared to the conventional tablets. Accordingly, the pH(M)-SD might be an efficient route for enhancing the dissolution and bioavailability of poorly water-soluble GT0918.


The immune response to sporadic colorectal cancer in a novel mouse model.
Current mouse models do not reflect the sporadic nature of colon cancer and do not allow the analysis of antitumor immune response because of the lack of known tumor-specific antigens. Two transgenic mouse models with spontaneous tumor development were generated, directing the expression of SV40T antigen (Tag) either constitutively (Vil-Cre × LoxP-Tag-transgenic mice) or stochastically (Vil-Cre-ER(T2) × LoxP-Tag-transgenic mice) into the putative stem cell region of the crypt of Lieberkühn. Tumor development and antitumor immune response were monitored. Vil-Cre × LoxP-Tag mice developed multiple adenocarcinomas of the small intestine and colon at an average age of 6 months. During the tumor development, Tag-specific immunoglobulin G (IgG) antibodies were induced in half of the mice, although they had developed neonatal cytotoxic T lymphocyte (CTL) tolerance. This model shows similarity to hereditary colon cancer but not to the sporadic tumor development. Therefore, the conditional Vil-Cre-ER(T2) × LoxP-Tag mice were established, in which expression of the dormant Tag was induced by stochastic, tissue-specific activation of Cre recombinase. These mice spontaneously developed highly invasive, metastasizing colon carcinomas at an average age of 20 months. Colon carcinomas expressed epithelial and/or neuroendocrine markers depending on the grade of differentiation. Young Vil-Cre-ER(T2) × LoxP-Tag mice had retained CTL responses against epitope IV of Tag. The tumors induced strong anti-Tag IgG responses. We report, for the first time, a mouse model based on stochastic, tissue-specific activation of a dormant oncogene in the colon allowing the analysis of antitumor immune response against primary colorectal cancer.


Infiltration of Solution-Processable Solid Electrolytes into Conventional Li-Ion-Battery Electrodes for All-Solid-State Li-Ion Batteries.
Bulk-type all-solid-state lithium-ion batteries (ASLBs) have the potential to be superior to conventional lithium-ion batteries (LIBs) in terms of safety and energy density. Sulfide SE materials are key to the development of bulk-type ASLBs because of their high ionic conductivity (max of ∼10-2 S cm-1) and deformability. However, the severe reactivity of sulfide materials toward common polar solvents and the particulate nature of these electrolytes pose serious complications for the wet-slurry process used to fabricate ASLB electrodes, such as the availability of solvent and polymeric binders and the formation of ionic contacts and networks. In this work, we report a new scalable fabrication protocol for ASLB electrodes using conventional composite LIB electrodes and homogeneous SE solutions (Li6PS5Cl (LPSCl) in ethanol or 0.4LiI-0.6Li4SnS4 in methanol). The liquefied LPSCl is infiltrated into the tortuous porous structures of LIB electrodes and solidified, providing intimate ionic contacts and favorable ionic percolation. The LPSCl-infiltrated LiCoO2 and graphite electrodes show high reversible capacities (141 and 364 mA h g-1) at 0.14 mA cm-2 (0.1 C) and 30 °C, which are not only superior to those for conventional dry-mixed and slurry-mixed ASLB electrodes but also comparable to those for liquid electrolyte cells. Good electrochemical performance of ASLBs employing the LPSCl-infiltrated LiCoO2 and graphite electrodes at 100 °C is also presented, highlighting the excellent thermal stability and safety of ASLBs.


Correlation of Echocardiogram and Exercise Test Data in Children with Aortic Stenosis.
Previous pediatric exercise test criteria for aortic stenosis severity were based on cardiac catheterization assessment, whereas current criteria are based on echocardiographic valve gradients. We sought to correlate exercise test criteria with echocardiographic assessment of severity. We report 65 studies, 51 patients (mean age of 13 ± 4 years; 75% males), with aortic stenosis (AS) who had a maximal exercise test between 2005 and 2016. We defined three groups based on resting mean Doppler gradient across their aortic valve: severe AS (n = 10; gradient of ≥ 40 mmHg), moderate AS (n = 20; gradient 25-39 mmHg), and mild AS (n = 35; gradient ≤ 24 mmHg). We studied symptoms (chest pain) during exercise, resting electrocardiogram changes (left ventricular hypertrophy [LVH]), complex arrhythmias during exercise, change in exercise systolic blood pressure (SBP; delta SBP = peak SBP-resting SBP), exercise duration, work, echocardiogram parameters (LVH), and ST-T wave changes with exercise. Additionally, we compared work and delta SBP during exercise with 117 control males and females without heart disease. Severe AS patients have statistically significant differences when compared with mild AS in ST-T wave depression during exercise, LVH on resting electrocardiogram, and echocardiogram. There was a significant difference in delta SBP between severe AS and normal controls (delta SBP 21.6 vs. 46.2 mmHg), and between moderate AS and normal controls (delta SBP 32 vs. 46.2 mmHg). There were no significant complications during maximal exercise testing. Children with echocardiographic severe and moderate AS have exercise testing abnormalities. Exercise test criteria for severity of AS were validated for echocardiographic criteria for AS severity.


Multiyear Factor VIII Expression after AAV Gene Transfer for Hemophilia A.
The goal of gene therapy for patients with hemophilia A is to safely impart long-term stable factor VIII expression that predictably ameliorates bleeding with the use of the lowest possible vector dose. In this phase 1-2 trial, we infused an investigational adeno-associated viral (AAV) vector (SPK-8011) for hepatocyte expression of factor VIII in 18 men with hemophilia A. Four dose cohorts were enrolled; the lowest-dose cohort received a dose of 5 × 1011 vector genomes (vg) per kilogram of body weight, and the highest-dose cohort received 2 × 1012 vg per kilogram. Some participants received glucocorticoids within 52 weeks after vector administration either to prevent or to treat a presumed AAV capsid immune response. Trial objectives included evaluation of the safety and preliminary efficacy of SPK-8011 and of the expression and durability of factor VIII. The median safety observation period was 36.6 months (range, 5.5 to 50.3). A total of 33 treatment-related adverse events occurred in 8 participants; 17 events were vector-related, including 1 serious adverse event, and 16 were glucocorticoid-related. Two participants lost all factor VIII expression because of an anti-AAV capsid cellular immune response that was not sensitive to immune suppression. In the remaining 16 participants, factor VIII expression was maintained; 12 of these participants were followed for more than 2 years, and a one-stage factor VIII assay showed no apparent decrease in factor VIII activity over time (mean [±SD] factor VIII activity, 12.9±6.9% of the normal value at 26 to 52 weeks when the participants were not receiving glucocorticoids vs. 12.0±7.1% of the normal value at >52 weeks after vector administration; 95% confidence interval [CI], -2.4 to 0.6 for the difference between matched pairs). The participants had a 91.5% reduction (95% CI, 88.8 to 94.1) in the annualized bleeding rate (median rate, 8.5 events per year [range, 0 to 43.0] before vector administration vs. 0.3 events per year [range, 0 to 6.5] after vector administration). Sustained factor VIII expression in 16 of 18 participants who received SPK-8011 permitted discontinuation of prophylaxis and a reduction in bleeding episodes. No major safety concerns were reported. (Funded by Spark Therapeutics and the National Heart, Lung, and Blood Institute; ClinicalTrials.gov numbers, NCT03003533 and NCT03432520.).


Validation of aortic valve pressure gradient quantification using semi-automated 4D flow CMR pipeline.
Doppler echocardiographic aortic valve peak velocity and peak pressure gradient assessment across the aortic valve (AV) is the mainstay for diagnosing aortic stenosis. Four-dimensional flow cardiovascular magnetic resonance (4D flow CMR) is emerging as a valuable diagnostic tool for estimating the peak pressure drop across the aortic valve, but assessment remains cumbersome. We aimed to validate a novel semi-automated pipeline 4D flow CMR method of assessing peak aortic value pressure gradient (AVPG) using the commercially available software solution, CAAS MR Solutions, against invasive angiographic methods. We enrolled 11 patients with severe AS on echocardiography from the EurValve programme. All patients had pre-intervention doppler echocardiography, invasive cardiac catheterisation with peak pressure drop assessment across the AV and 4D flow CMR. The peak AVPG was 51.9 ± 35.2 mmHg using the invasive pressure drop method and 52.2 ± 29.2 mmHg for the 4D flow CMR method (semi-automated pipeline), with good correlation between the two methods (r = 0.70, p = 0.017). Assessment of AVPG by 4D flow CMR using the novel semi-automated pipeline method shows excellent agreement to invasive assessment when compared to doppler-based methods and advocate for its use as complementary to echocardiography.


Tracheobronchial mucus: abnormalities related to asthma and its treatment.
The presence of mucociliary dysfunction in bronchial asthma is suggested by clinical observations and has been confirmed by pathologic and physiologic studies. Since mucociliary dysfunction may play a considerable role in the pathophysiology of bronchial asthma, currently available therapeutic measures which enhance mucous transport should be included in the treatment plan. It is expected that a better understanding of the underlying mechanisms will ultimately result in the development of more potent pharmacologic agents capable of improving abnormal mucociliary function.


The N-Terminal Flanking Region Modulates the Actin Binding Affinity of the Utrophin Tandem Calponin-Homology Domain.
Despite sharing a high degree of sequence similarity, the tandem calponin-homology (CH) domain of utrophin binds to actin 30 times stronger than that of dystrophin. We have previously shown that this difference in actin binding affinity could not be ascribed to the differences in inter-CH-domain linkers [Bandi, S., et al. (2015) Biochemistry 54, 5480-5488]. Here, we examined the role of the N-terminal flanking region. The utrophin tandem CH domain contains a 27-residue flanking region before its CH1 domain. We examined its effect by comparing the structure and function of full-length utrophin tandem CH domain Utr(1-261) and its truncated Utr(28-261) construct. Both full-length and truncated constructs are monomers in solution, with no significant differences in their secondary or tertiary structures. Truncated construct Utr(28-261) binds to actin 30 times weaker than that of the full-length Utr(1-261), similar to that of the dystrophin tandem CH domain with a much shorter flanking region. Deletion of the N-terminal flanking region stabilizes the CH1 domain. The magnitude of the change in binding free energy upon truncation is similar to that of the change in thermodynamic stability. The isolated N-terminal peptide by itself is significantly random coil and does not bind to F-actin in the affinity range of Utr(1-261) and Utr(28-261). These results indicate that the N-terminal flanking region significantly affects the actin binding affinity of tandem CH domains. This observation further stresses that protein regions other than the three actin-binding surfaces identified earlier, irrespective of whether they directly bind to actin, also contribute to the actin binding affinity of tandem CH domains.


Does the bracket-ligature combination affect the amount of orthodontic space closure over three months? A randomized controlled trial.
To investigate the effect of bracket-ligature combination on the amount of orthodontic space closure over three months. Randomized clinical trial with three parallel groups. A hospital orthodontic department (Chesterfield Royal Hospital, UK). Forty-five patients requiring upper first premolar extractions. Informed consent was obtained and participants were randomly allocated into one of three groups: (1) conventional pre-adjusted edgewise brackets and elastomeric ligatures; (2) conventional pre-adjusted edgewise brackets and Super Slick(®) low friction elastomeric ligatures; (3) Damon 3MX(®) passive self-ligating brackets. Space closure was undertaken on 0·019×0·025-inch stainless steel archwires with nickel-titanium coil springs. Participants were recalled at four weekly intervals. Upper alginate impressions were taken at each visit (maximum three). The primary outcome measure was the mean amount of space closure in a 3-month period. A one-way ANOVA was undertaken [dependent variable: mean space closure (mm); independent variable: group allocation]. The amount of space closure was very similar between the three groups (1 mm per 28 days); however, there was a wide variation in the rate of space closure between individuals. The differences in the amount of space closure over three months between the three groups was very small and non-significant (P = 0·718). The hypothesis that reducing friction by modifying the bracket/ligature interface increases the rate of space closure was not supported. The major determinant of orthodontic tooth movement is probably the individual patient response.


The Future of Cesarean Delivery Rates in the United States.
The US cesarean delivery rate remains 30%-32%. Increases in maternal age, obesity, and diabetes put upward pressure on this rate. Alternatives to cesarean delivery, vaginal birth after cesarean (VBAC), and operative vaginal delivery, are underutilized and there are substantial challenges to their resurgence. Practice guidelines offer promise, but demonstrate only minor reductions in cesarean delivery. We estimate that the overall rate in the US will remain 27%-30% for the immediate future. As more states move to recognize the independent practice of midwifery and more payers seek lower cost options for childbirth, we anticipate the overall rate will drop to 20% or 25%, but not for another 15 years or more.


Appendicular skeletal muscle in hospitalised hip-fracture patients: development and cross-validation of anthropometric prediction equations against dual-energy X-ray absorptiometry.
accurate and practical assessment methods for assessing appendicular skeletal muscle (ASM) is of clinical importance for the diagnosis of geriatric syndromes associated with skeletal muscle wasting. the purpose of this study was to develop and cross-validate novel anthropometric prediction equations for the estimate of ASM in older adults post-surgical fixation for hip fracture, using dual-energy X-ray absorptiometry (DEXA) as the criterion measure. community-dwelling older adults (aged ≥65 years) recently hospitalised for hip fracture. participants were recruited from hospital in the acute phase of recovery. validation measurement study. a total of 79 hip fracture patients were involved in the development of the regression models (MD group). A further 64 hip fracture patients also recruited in the early phase of recovery were used in the cross-validation of the regression models (CV group). Multiple linear regression analyses were undertaken in the MD group to identify the best performing prediction models. The linear coefficient of determination (R(2)) in addition to the standard error of the estimate (SEE) were calculated to determine the best performing model. Agreement between estimated ASM and ASMDEXA in the CV group was assessed using paired t-tests with the 95% limits of agreement (LOA) assessed using Bland-Altman analyses. the mean age of all the participants was 82.1 ± 7.3 years. The best two prediction models are presented as follows: ASMPRED-EQUATION_1: 22.28 - (0.069 * age) + (0.407 * weight) - (0.807 * BMI) - (0.222 * MAC) (adjusted R(2): 0.76; SEE: 1.80 kg); ASMPRED-EQUATION_2: 16.77 - (0.036 * age) + (0.385 * weight) - (0.873 * BMI) (adjusted R(2): 0.73; SEE: 1.90 kg). The mean bias from the CV group between ASMDEXA and the predictive equations is as follows: ASMDEXA - ASMPRED-EQUATION_1: 0.29 ± 2.6 kg (LOA: -4.80, 5.40 kg); ASMDEXA - ASMPRED-EQUATION_2: 0.13 ± 2.5 kg (LOA: -4.77, 5.0 kg). No significant difference was observed between measured ASMDEXA and estimated ASM (ASMDEXA: 16.4 ± 3.9 kg; ASMPRED-EQUATION_1: 16.7 ± 3.2 kg (P = 0.379); ASMPRED-EQUATION_2: 16.6 ± 3.2 kg (P = 0.670)). we have developed and cross-validated novel anthropometric prediction equations against DEXA for the estimate of ASM designed for application in older orthopaedic patients. Our equation may be of use as an alternative to DEXA in the diagnosis of skeletal muscle wasting syndromes. Further validation studies are required to determine the clinical utility of our equation across other settings, including hip fracture patients admitted from residential care, and also with a longer-term follow-up.


Signals through gp130 upregulate bcl-x gene expression via STAT1-binding cis-element in cardiac myocytes.
We described recently the activation of the Janus kinasesignal transducer and activator of transcription (JakSTAT) and mitogen-activated protein (MAP) kinase pathways by leukemia inhibitory factor (LIF) through gp130, a signal transducer of IL-6-related cytokines, that transduces hypertrophic signals in cardiac myocytes. In addition, stimulation of gp130 by IL-6-related cytokines is known to exert a cytoprotective effect. In the present study, we investigated the possibility that activation of gp130 initiates activation of the cytoprotective genes in cardiac myocytes. Incubation of cardiac myocytes with LIF induced the expression of bcl-x, and the isoform that was induced by LIF was identified as bcl-xL. Induction of bcl-xL protein was also identified by Western blotting. Antisense oligonucleotide against bcl-x mRNA inhibited protective effect of LIF accompanied with the reduction in bclxL protein. We constructed bcl-x promoter-luciferase reporter gene plasmids (-639/+10- or -161/+10-luciferase), and transfected them to cardiac myocytes. LIF stimulation increased the luciferase activity of -639/+10-luciferase plasmids. Although -161/+10-luciferase plasmids presented comparable responsiveness to LIF, the basal transcription level was impaired. The LIF-responsive cis-element was localized to a DNA fragment (positions -161 to +10) that contains an interferon-gamma activation site (GAS) motif (GGA) at position -41 of the bcl-x gene promoter. This motif bound to STAT1, not to STAT3, and site-directed mutagenesis revealed that this motif was essential for LIF-responsive promoter activity. These data suggest that LIF induces bcl-x mRNA via STAT1 binding cis-element in cardiac myocytes, presenting cytoprotective effect.


Differential phosphorylation of (E)-5-(2-bromovinyl)-2'-deoxyuridine monophosphate by thymidylate kinases from herpes simplex viruses types 1 and 2 and varicella zoster virus.
5-(2-Bromovinyl)-2'-deoxyuridine (BrVdUrd) is a potent and selective inhibitor of herpes simplex virus Type I (HS-I) and varicella zoster (VZ) virus replication but is much less potent against herpes simplex virus Type II (HS-II) replication. A possible enzymatic basis for this difference is reported here to involve the virus-coded dThd-dTMP kinases (EC 220.127.116.11) from the three virus strains. The thymidine kinases from the three virus strains were purified by affinity chromatography. In addition to catalyzing the phosphorylation of nucleosides, each of the three purified enzymes catalyzed the phosphorylation of thymidylate to its diphosphate but at strikingly different rates. The relative amounts of extractable virus-coded thymidylate kinases were estimated to be 100/2/40 for cells infected with HS-I, HS-II, and VZ viruses, respectively. Extracts of cells infected with HS-I virus catalyzed the phosphorylation of the monophosphate of BrVdUrd to its diphosphate. In contrast, the product was not detected with extracts from cells infected with HS-II virus. THe ratios of rates with 0.5 mM BrVdUrd monophosphate versus 0.1 mM dTMP as substrates for each of the purified dThd-dTMP kinases from HS-I, HS-II, and VZ viruses and the dTMP kinase from host cells were, respectively, 0.09, less than 0.002, 0.03, and less than 0.0002. These observations correlate with the relative sensitivities of these viruses to BrVdUrd in cell culture and suggest that, if BrVdUrd exerts its effect as a triphosphate, the inefficient phosphorylation of the monophosphate contributes to the insensitivity of HS-II virus to this agent.


Magnetization vector manipulation by electric fields.
Conventional semiconductor devices use electric fields to control conductivity, a scalar quantity, for information processing. In magnetic materials, the direction of magnetization, a vector quantity, is of fundamental importance. In magnetic data storage, magnetization is manipulated with a current-generated magnetic field (Oersted-Ampère field), and spin current is being studied for use in non-volatile magnetic memories. To make control of magnetization fully compatible with semiconductor devices, it is highly desirable to control magnetization using electric fields. Conventionally, this is achieved by means of magnetostriction produced by mechanically generated strain through the use of piezoelectricity. Multiferroics have been widely studied in an alternative approach where ferroelectricity is combined with ferromagnetism. Magnetic-field control of electric polarization has been reported in these multiferroics using the magnetoelectric effect, but the inverse effect-direct electrical control of magnetization-has not so far been observed. Here we show that the manipulation of magnetization can be achieved solely by electric fields in a ferromagnetic semiconductor, (Ga,Mn)As. The magnetic anisotropy, which determines the magnetization direction, depends on the charge carrier (hole) concentration in (Ga,Mn)As. By applying an electric field using a metal-insulator-semiconductor structure, the hole concentration and, thereby, the magnetic anisotropy can be controlled, allowing manipulation of the magnetization direction.


Adenovirus encoding human platelet-derived growth factor-B delivered to alveolar bone defects exhibits safety and biodistribution profiles favorable for clinical use.
Platelet-derived growth factor (PDGF) gene therapy offers promise for tissue engineering of tooth-supporting alveolar bone defects. To date, limited information exists regarding the safety profile and systemic biodistribution of PDGF gene therapy vectors when delivered locally to periodontal osseous defects. The aim of this preclinical study was to determine the safety profile of adenovirus encoding the PDGF-B gene (AdPDGF-B) delivered in a collagen matrix to periodontal lesions. Standardized alveolar bone defects were created in rats, followed by delivery of matrix alone or containing AdPDGF-B at 5.5 x 10(8) or 5.5 x 10(9) plaque-forming units/ml. The regenerative response was confirmed histologically. Gross clinical observations, hematology, and blood chemistries were monitored to evaluate systemic involvement. Bioluminescence and quantitative polymerase chain reaction were used to assess vector biodistribution. No significant histopathological changes were noted during the investigation. Minor alterations in specific hematological and blood chemistries were seen; however, most parameters were within the normal range for all groups. Bioluminescence analysis revealed vector distribution at the axillary lymph nodes during the first 2 weeks with subsequent return to baseline levels. AdPDGF-B was well contained within the localized osseous defect area without viremia or distant organ involvement. These results indicate that AdPDGF-B delivered in a collagen matrix exhibits acceptable safety profiles for possible use in human clinical studies.


Spectrophotometry of carnitine in biological fluids and tissue with a Cobas Bio centrifugal analyzer.
A spectrophotometric method for carnitine has been adapted to the Cobas Bio centrifugal analyzer. The addition of carnitine to a system containing carnitine acetyltransferase (EC 22.214.171.124) and acetyl-CoA gives rise to the formation of CoA. The system is coupled to 5,5'-dithiobis(2-nitrobenzoate) (DTNB). Assay response varied linearly with concentration of carnitine over a wide concentration range. The total CV was 5.5% for a carnitine concentration in serum of 58.0 mumol/L. Analytical recovery of carnitine added to a serum sample was 93%. No interference was found in icteric, not grossly hemolyzed, lipemic, or uremic sera. Comparison with a radioenzymatic method showed that results correlated well (r greater than 0.965) but the present method gave values proportionally greater by 10 to 25% for samples of plasma, dialysis fluid, urine, and muscle tissue. Advantages over the original spectrophotometric assays involving DTNB include low reagent costs, rapidity, simplicity, and reproducibility. However, this modification is not as sensitive and probably not as specific as the radioenzymatic methods.


Mouse and human CRKL is dosage sensitive for cardiac outflow tract formation.
The human chromosome 22q11.2 region is susceptible to rearrangements during meiosis leading to velo-cardio-facial/DiGeorge/22q11.2 deletion syndrome (22q11DS) characterized by conotruncal heart defects (CTDs) and other congenital anomalies. The majority of individuals have a 3 Mb deletion whose proximal region contains the presumed disease-associated gene TBX1 (T-box 1). Although a small subset have proximal nested deletions including TBX1, individuals with distal deletions that exclude TBX1 have also been identified. The deletions are flanked by low-copy repeats (LCR22A, B, C, D). We describe cardiac phenotypes in 25 individuals with atypical distal nested deletions within the 3 Mb region that do not include TBX1 including 20 with LCR22B to LCR22D deletions and 5 with nested LCR22C to LCR22D deletions. Together with previous reports, 12 of 37 (32%) with LCR22B-D deletions and 5 of 34 (15%) individuals with LCR22C-D deletions had CTDs including tetralogy of Fallot. In the absence of TBX1, we hypothesized that CRKL (Crk-like), mapping to the LCR22C-D region, might contribute to the cardiac phenotype in these individuals. We created an allelic series in mice of Crkl, including a hypomorphic allele, to test for gene expression effects on phenotype. We found that the spectrum of heart defects depends on Crkl expression, occurring with analogous malformations to that in human individuals, suggesting that haploinsufficiency of CRKL could be responsible for the etiology of CTDs in individuals with nested distal deletions and might act as a genetic modifier of individuals with the typical 3 Mb deletion.


Interaction between Bovine Serum Albumin in Fresh Milk Cream and Encapsulated and Non-Encapsulated Polyphenols of Tamarillo.
The fortification of dairy products with polyphenols is known to deliver additional health benefits. However, interactions between polyphenols may form complexes and cause a loss of functionality overall. This study aimed to investigate potential interactions between polyphenols, in encapsulated and non-encapsulated forms, extracted from tamarillo fruit and bovine serum albumin (BSA) from fresh milk cream. Fortification with tamarillo extract was made at 1, 2 and 3% (w/w), and the resultant changes in physicochemical, rheological and functional properties were studied. With an increase in fortification, the absorbance of protein-ligand in the protein-polyphenol complex was decreased by up to 55% and 67% in UV and fluorescent intensities, respectively. Chlorogenic acid and kaempferol-3-rutinoside were more affected than delphinidin-3-rutinoside and pelargonidin-3-rutinoside. Static quenching was the main mechanism in the fluorescence spectra. Tryptophan and tyrosine residues were the two major aromatic amino acids responsible for the interactions with BSA. There were at least three binding sites near the tryptophan residue on BSA. The rheological property remained unaffected after the addition of non-encapsulated tamarillo extracts. Antioxidant capacity was significantly decreased (p < 0.05) after the addition of encapsulated extracts. This may be explained by using a low concentration of maltodextrin (10% w/w) as an encapsulating agent and its high binding affinity to milk proteins.


Good farming practices in apiculture.
Modern European beekeeping is facing numerous challenges due to a variety of factors, mainly related to globalisation, agrochemical pollution and environmental changes. In addition to this, new pathogens threaten the health of European honey bees. In that context, correct colony management should encompass a wider vision, where productivity aspects are linked to a One Health approach in order to protect honey bees, humans and the environment. This paper describes a novel tool to be applied in beekeeping operations: good beekeeping practices (GBPs). The authors ranked a list of GBPs scored against their importance and validated by an international team, including researchers, national animal health authorities and international beekeepers' associations. These activities were carried out in the project 'BPRACTICES', approved within the transnational call of the European Research Area Network on Sustainable Animal Production (ERA-NET SusAn) in the Horizon 2020 Research and Innovation Programme of the European Union. This study, created through an international collaboration, aims to present an innovative and implementable approach, similar to applications already adopted in other livestock production systems.


Delineating transitions during the evolution of specialised peroxisomes: Glycosome formation in kinetoplastid and diplonemid protists.
One peculiarity of protists belonging to classes Kinetoplastea and Diplonemea within the phylum Euglenozoa is compartmentalisation of most glycolytic enzymes within peroxisomes that are hence called glycosomes. This pathway is not sequestered in peroxisomes of the third Euglenozoan class, Euglenida. Previous analysis of well-studied kinetoplastids, the 'TriTryps' parasites Trypanosoma brucei, Trypanosoma cruzi and Leishmania spp., identified within glycosomes other metabolic processes usually not present in peroxisomes. In addition, trypanosomatid peroxins, i.e. proteins involved in biogenesis of these organelles, are divergent from human and yeast orthologues. In recent years, genomes, transcriptomes and proteomes for a variety of euglenozoans have become available. Here, we track the possible evolution of glycosomes by querying these databases, as well as the genome of Naegleria gruberi, a non-euglenozoan, which belongs to the same protist supergroup Discoba. We searched for orthologues of TriTryps proteins involved in glycosomal metabolism and biogenesis. Predicted cellular location(s) of each metabolic enzyme identified was inferred from presence or absence of peroxisomal-targeting signals. Combined with a survey of relevant literature, we refine extensively our previously postulated hypothesis about glycosome evolution. The data agree glycolysis was compartmentalised in a common ancestor of the kinetoplastids and diplonemids, yet additionally indicates most other processes found in glycosomes of extant trypanosomatids, but not in peroxisomes of other eukaryotes were either sequestered in this ancestor or shortly after separation of the two lineages. In contrast, peroxin divergence is evident in all euglenozoans. Following their gain of pathway complexity, subsequent evolution of peroxisome/glycosome function is complex. We hypothesize compartmentalisation in glycosomes of glycolytic enzymes, their cofactors and subsequently other metabolic enzymes provided selective advantage to kinetoplastids and diplonemids during their evolution in changing marine environments. We contend two specific properties derived from the ancestral peroxisomes were key: existence of nonselective pores for small solutes and the possibility of high turnover by pexophagy. Critically, such pores and pexophagy are characterised in extant trypanosomatids. Increasing amenability of free-living kinetoplastids and recently isolated diplonemids to experimental study means our hypothesis and interpretation of bioinformatic data are suited to experimental interrogation.


The call of the wild: using non-model systems to investigate microbiome-behaviour relationships.
On and within most sites across an animal's body live complex communities of microorganisms. These microorganisms perform a variety of important functions for their hosts, including communicating with the brain, immune system and endocrine axes to mediate physiological processes and affect individual behaviour. Microbiome research has primarily focused on the functions of the microbiome within the gastrointestinal tract (gut microbiome) using biomedically relevant laboratory species (i.e. model organisms). These studies have identified important connections between the gut microbiome and host immune, neuroendocrine and nervous systems, as well as how these connections, in turn, influence host behaviour and health. Recently, the field has expanded beyond traditional model systems as it has become apparent that the microbiome can drive differences in behaviour and diet, play a fundamental role in host fitness and influence community-scale dynamics in wild populations. In this Review, we highlight the value of conducting hypothesis-driven research in non-model organisms and the benefits of a comparative approach that assesses patterns across different species or taxa. Using social behaviour as an intellectual framework, we review the bidirectional relationship between the gut microbiome and host behaviour, and identify understudied mechanisms by which these effects may be mediated.


The protective effects of self-compassion on alcohol-related problems among First Nation adolescents.
Given the disproportionate alcohol-related consequences experienced by North American Indigenous youth, there is a critical need to identify related risk and protective factors. Self-compassion, which has been found to mitigate the effects of trauma exposure, may serve as one such protective factor given the high-degree of historical trauma and contemporary discrimination identified as contributing to the alcohol-related disparities experienced by Indigenous communities. However, no research has examined how self-compassion (i.e., the ability to be kind and accepting and to extend compassion towards oneself) plays a unique role in Indigenous peoples' experiences with alcohol. First Nation adolescents between the ages of 11 and 18 living on a reserve in Eastern Canada (N=106, M age =14.6, 50.0% female) completed a pencil-and-paper survey regarding their alcohol use, alcohol-related problems, and self-compassion. Self-compassion was significantly inversely associated with alcohol-related problems (b=-.51, p=.01, 95%CI [-.90, -.12], and significantly interacted with frequency of alcohol use in predicting alcohol-related problems (b=-.42, p=.04, 95%CI [-.82, -.03]). Simple slopes analyses revealed that the association between frequency of alcohol use and frequency of experiencing alcohol-related problems was significant and positive at low (b=4.68, p<.001, 95%CI [2.62, 6.73]), but was not significant at high (b=-.29, p=.89, 95%CI [-4.35, 3.77]) levels of self-compassion. Binary logistic regression revealed that higher scores of self-compassion were associated with a lower odds of being in the high-risk group for AUD (OR=0.90, 95%CI [0.83, 0.98], p=.02). Our results suggest self-compassion may be protective against experiencing alcohol problems in Indigenous youth and thus may be a target for behavioral interventions.


PAS domain containing regulator SLCG_7083 involved in morphological development and glucose utilization in Streptomyces lincolnensis.
Streptomyces lincolnensis is well known for producing the clinically important antimicrobial agent lincomycin. The synthetic and regulatory mechanisms on lincomycin biosynthesis have been deeply explored in recent years. However, the regulation involved in primary metabolism have not been fully addressed. SLCG_7083 protein contains a Per-Arnt-Sim (PAS) domain at the N-terminus, whose homologous proteins are highly distributed in Streptomyces. The inactivation of the SLCG_7083 gene indicated that SLCG_7083 promotes glucose utilization, slows mycelial growth and affects sporulation in S. lincolnensis. Comparative transcriptomic analysis further revealed that SLCG_7083 represses eight genes involved in sporulation, cell division and lipid metabolism, and activates two genes involved in carbon metabolism. SLCG_7083 is a PAS domain-containing regulator on morphological development and glucose utilization in S. lincolnensis. Our results first revealed the regulatory function of SLCG_7083, and shed new light on the transcriptional effects of SLCG_7083-like family proteins in Streptomyces.


Lab on a chip for multiplexed immunoassays to detect bladder cancer using multifunctional dielectrophoretic manipulations.
A multiplexed immunosensor has been developed for the detection of specific biomarkers Galectin-1 (Gal-1) and Lactate Dehydrogenase B (LDH-B) present in different grades of bladder cancer cell lysates. In order to immobilize nanoprobes with different antibodies on a single chip we employed three-step programmable dielectrophoretic manipulations for focusing, guiding and trapping to enhance the fluorescent response and reduce the interference between the two antibody arrays. The chip consisted of a patterned indium tin oxide (ITO) electrode for sensing and a middle fish bone shaped gold electrode for focusing and guiding. Using ITO electrodes for the sensing area can effectively eliminate the background noise of fluorescence response as compared to metal electrodes. It was also observed that the three step manipulation increased fluorescence response after immunosensing by about 4.6 times as compared to utilizing DEP for just trapping the nanoprobes. Two different-grade bladder cancer cell lysates (grade I: RT4 and grade III: T24) were individually analyzed for detecting the protein expression levels of Gal-1 and LDH-B. The fluorescence intensity observed for Gal-1 is higher than that of LDH-B in the T24 cell lysate; however the response observed in RT4 is higher for LDH-B as compared to Gal-1. Thus we can effectively identify the different grades of bladder cancer cells. In addition, the platform for DEP manipulation developed in this study can enable real time detection of multiple analytes on a single chip and provide more practical benefits for clinical diagnosis.


Comparative study of conservative treatment and lymphaticovenular anastomosis with compression therapy for early-stage breast cancer-related lymphoedema.
This single-centre retrospective cohort study aimed to clarify the superiority of treatment by comparing the outcomes of lymphaticovenular anastomosis with compression therapy and conservative treatment centred on compression therapy in the early stage of breast cancer-related lymphoedema. Data were collected from all patients treated for breast cancer-related lymphoedema between January 2015 and December 2022. The patients were classified into conservative treatment and surgical treatment groups. The upper extremity lymphoedema index value was calculated, based on five circumference values of the upper extremity and body mass index, to compare the 6-, 12-, 18- and 24-month outcomes between the groups. Of 101 patients with breast cancer-related lymphoedema, 81 (conservative treatment: 52; surgical treatment: 29) were included in the analysis. The therapeutic effect was significantly higher in the surgical treatment group than in the conservative treatment group, when comparing the rate of change in oedema at 6 (-6.6% ± 7.3% vs. 0.9% ± 7.5%; p < 0.001), 12 (-7.3% ± 6.2% vs. 2.9% ± 8.6%; p < 0.001), 18 (-7.6% ± 8.0% vs. 3.9% ± 9.2%; p < 0.001) and 24 (-5.6% ± 6.0% vs. 4.4% ± 10.7%; p < 0.001) months. The incidence of cellulitis increased in the conservative treatment group (from 9.6% to 15.4%), whereas it was suppressed in the surgical treatment group (from 13.8% to 0%). Conservative treatment centred on compression therapy increased oedema over time; however, lymphaticovenular anastomosis with compression therapy effectively reduced oedema.


Vagal regulation of heart rate in the prediction of developmental outcome for very low birth weight preterm infants.
To investigate heart rate and respiratory sinus arrhythmia (RSA) as markers of developmental outcome, infant ECG and 3 year outcome were assessed in 41 very low birth weight (< 1,500 g) infants. Measures of mean heart rate and RSA, and the maturational shifts in their values from 33 to 35 weeks gestational age, were recorded. RSA measures predicted 3 year outcome beyond the effects of birth weight, medical risk, and socioeconomic status. Higher RSA was associated with better social skills, whereas greater RSA maturation was associated with better mental processing and gross motor skills. Lower heart rate was associated with better behavior regulation and social skills, whereas greater maturational decreases were associated with better gross motor skills. Dividing the sample into groups of infants with birth weights less than 1,000 g and those with birth weights over 1,000 g, RSA maturation emerged a strong predictor of mental processing, knowledge base, and gross motor skills in the former. A measure of joint maturation of RSA and heart rate was associated with better behavior regulation at 3 years, as measured by Child Behavior Checklist and Parenting Stress Index scores, for this group. The findings directly respond to the need for physiological variables in the prediction of outcome in high-risk infants.


The women on Stieve's list: Victims of national socialism whose bodies were used for anatomical research.
Research on the history of anatomy in the Third Reich has often concentrated on the influence of the National Socialist (NS) regime on anatomists and their consequent unethical activities. Only recently, the focus has shifted to NS victims whose bodies were used for anatomical purposes. As a first approach to learning more about the victims, this study investigated the persons whose names Hermann Stieve, chairman of the Anatomical Department at the University of Berlin, had listed after using their bodies for his research. The study draws a group portrait and recounts selected biographies of the 174 women and eight men on the list. Most women were of reproductive age, two-thirds were German and a majority was executed for political reasons. Among the executed were at least two pregnant women. The corrected names, biographical data, and nationalities of all persons on the list are published here. None of them volunteered to be dissected, nor were the anatomists at the time interested in the victims' personal background. Future work will have to focus on the investigation of further biographies so that numbers can be turned back into people. This history is a reminder to modern anatomy that ethical body procurement and the anatomists' caring about the body donor is of the utmost importance in a discipline that introduces students to professional ethics in the medical teaching curriculum.


Targeted Temperature Management: Quantifying the Extent of Serum Electrolyte and Blood Glucose Shifts in Postcardiac Arrest Patients.
This study aims to quantify the extent of electrolyte (potassium, magnesium, and phosphorus) and blood glucose changes during targeted temperature management (TTM), with insight on electrolyte replacements and insulin administration. This was a 3-year retrospective study of patients who underwent TTM via Arctic Sun. Electrolyte and blood glucose values in addition to electrolyte replacements and insulin administrations were collected before, during, and post-TTM. The primary analysis assessed changes in electrolyte and blood glucose values during and after TTM, and the secondary analysis assessed changes before and during, and before and after TTM. The secondary analysis also incorporated amount of electrolyte replacements and insulin administrations patients received before, during, and post-TTM. Sixty patients were included for analysis. Comparing levels during to after TTM, there was a significant increase in potassium (3.7 [0.7]-4.4 [0.7] mmol/L, p < 0.001) and decrease in blood glucose (192 [135]-134 [55] mg/dL, p = 0.001). Comparing levels before to during TTM, there was a significant decrease in potassium (4.3 [0.7]-3.7 [0.7] mmol/L, p < 0.001) and phosphorus (4.8 [3.2]-3.4 [1.5] mg/dL, p = 0.003). Comparing levels before to after TTM, there was a significant decrease in phosphorus (4.5 [2.9]-3.3 [1.2] mmol/L, p = 0.026) and blood glucose (219 [35]-134 [55] mg/dL, p < 0.001). Patients received on average potassium 102 mEq, magnesium 1.9 g, phosphorus 9 mmol, and insulin 94 units. Potassium significantly decreased during and significantly increased after TTM. Phosphorus significantly decreased during TTM and blood glucose significantly decreased after TTM. There were no significant changes in magnesium during the defined time period.


Antibacterial Activity and Pharmacokinetic Profile of a Promising Antibacterial Agent: 22-(2-Amino-phenylsulfanyl)-22-Deoxypleuromutilin.
A new pleuromutilin derivative, 22-(2-amino-phenylsulfanyl)-22-deoxypleuromutilin (amphenmulin), has been synthesized and proved excellent in vitro and in vivo efficacy than that of tiamulin against methicillin-resistant Staphylococcus aureus (MRSA), suggesting this compound may lead to a promising antibacterial agent to treat MRSA infections. In this study, the effectiveness and safety of amphenmulin were further investigated. Amphenmulin showed excellent antibacterial activity against MRSA (minimal inhibitory concentration = 0.0156~8 µg/mL) and performed time-dependent growth inhibition and a concentration-dependent postantibiotic effect (PAE). Acute oral toxicity test in mice showed that amphenmulin was a practical non-toxic drug and possessed high security as a new drug with the 50% lethal dose (LD50) above 5000 mg/kg. The pharmacokinetic properties of amphenmulin were then measured. After intravenous administration, the elimination half-life (T1/2), total body clearance (Clβ), and area under curve to infinite time (AUC0→∞) were 1.92 ± 0.28 h, 0.82 ± 0.09 L/h/kg, and 12.23 ± 1.35 μg·h/mL, respectively. After intraperitoneal administration, the T1/2, Clβ/F and AUC0→∞ were 2.64 ± 0.72 h, 4.08 ± 1.14 L/h/kg, and 2.52 ± 0.81 μg·h/mL, respectively, while for the oral route were 2.91 ± 0.81 h, 6.31 ± 2.26 L/h/kg, 1.67 ± 0.66 μg·h/mL, respectively. Furthermore, we evaluated the antimicrobial activity of amphenmulin in an experimental model of MRSA wound infection. Amphenmulin enhanced wound closure and promoted the healing of wound, which inhibited MRSA bacterial counts in the wound and decreased serum levels of the pro-inflammatory cytokines TNF-α, IL-6, and MCP-1.


Comparing gatifloxacin and clarithromycin in pneumonia symptom resolution and process of care.
In looking for outcome differences beyond rates of cure, we prospectively compared the symptom resolution, side effects, and processes of care between the use of clarithromycin and gatifloxacin for the treatment of radiographically confirmed community-acquired pneumonia. We conducted a multicenter, randomized, open-label study comparing gatifloxacin monotherapy to clarithromycin alone or combined with ceftriaxone for patients with multiple risk factors. We measured the return to usual activities and symptoms over seven interviews ending 42 days after randomization. Admission and hospital discharge decision support were provided to treating physicians. We enrolled 266 patients over the age of 18 years between September 2000 and June 2003. The groups were similar in age and gender, with a mean age of 53.5+/-19.4 years, and were 54% female. Patient severity as determined by the number of risk factors and the Pneumonia Severity Index was similar between groups; 95% of the patients were low risk. A total of 91% of patients completed at least five of seven symptom interviews. In the clarithromycin study arm, 64% received concomitant therapy with ceftriaxone. We found no significant difference in return to usual activities, pneumonia-specific symptom scores, and 12-item short-form health survey scores. Individual symptom scores were similar except for bad taste and injection site soreness, which were higher in clarithromycin patients. The rates of hospital admission and length of stay were similar. The cost of antibiotic was higher in the clarithromycin group: $257 versus $110 for gatifloxacin. We found that gatifloxacin monotherapy is similar to clarithromycin given with or without ceftriaxone for the treatment of community-acquired pneumonia, except that antibiotic cost, bad taste, and injection site soreness favor the use of gatifloxacin.


Competition in the pharmaceutical industry: how do quality differences shape advertising strategies?
We present a Hotelling model of price and advertising competition between prescription drugs that differ in quality/side effects. Promotional effort results in the endogenous formation of two consumer groups: brand loyal and non-brand loyal ones. We show that advertising intensities are strategic substitutes, with the better quality drugs being the ones that are most advertised. This positive association stems from the higher rents that firms can extract from consumers whose brand loyalty is endogenously determined by promotional effort. The model's main results on advertising and pricing strategies are taken to the data. The latter consists of product level data on prices and quantities, product level advertising data, as well as the qualitative information on drug quality contained in the Orange Book compiled by the Food and Drug Administration (FDA). The empirical results provide strong support to the model's predictions.


Methylotrophs and Methylotroph Populations for Chloromethane Degradation.
Chloromethane is a halogenated volatile organic compound, produced in large quantities by terrestrial vegetation. After its release to the troposphere and transport to the stratosphere, its photolysis contributes to the degradation of stratospheric ozone. A better knowledge of chloromethane sources (production) and sinks (degradation) is a prerequisite to estimate its atmospheric budget in the context of global warming. The degradation of chloromethane by methylotrophic communities in terrestrial environments is a major underestimated chloromethane sink. Methylotrophs isolated from soils, marine environments and more recently from the phyllosphere have been grown under laboratory conditions using chloromethane as the sole carbon source. In addition to anaerobes that degrade chloromethane, the majority of cultivated strains were isolated in aerobiosis for their ability to use chloromethane as sole carbon and energy source. Among those, the Proteobacterium Methylobacterium (recently reclassified as Methylorubrum) harbours the only characterisized 'chloromethane utilization' (cmu) pathway, so far. This pathway is not representative of chloromethane-utilizing populations in the environment as cmu genes are rare in metagenomes. Recently, combined 'omics' biological approaches with chloromethane carbon and hydrogen stable isotope fractionation measurements in microcosms, indicated that microorganisms in soils and the phyllosphere (plant aerial parts) represent major sinks of chloromethane in contrast to more recently recognized microbe-inhabited environments, such as clouds. Cultivated chloromethane-degraders lacking the cmu genes display a singular isotope fractionation signature of chloromethane. Moreover, 13CH3Cl labelling of active methylotrophic communities by stable isotope probing in soils identify taxa that differ from the taxa known for chloromethane degradation. These observations suggest that new biomarkers for detecting active microbial chloromethane-utilizers in the environment are needed to assess the contribution of microorganisms to the global chloromethane cycle.


Cross-organism learning method to discover new gene functionalities.
Knowledge of gene and protein functions is paramount for the understanding of physiological and pathological biological processes, as well as in the development of new drugs and therapies. Analyses for biomedical knowledge discovery greatly benefit from the availability of gene and protein functional feature descriptions expressed through controlled terminologies and ontologies, i.e., of gene and protein biomedical controlled annotations. In the last years, several databases of such annotations have become available; yet, these valuable annotations are incomplete, include errors and only some of them represent highly reliable human curated information. Computational techniques able to reliably predict new gene or protein annotations with an associated likelihood value are thus paramount. Here, we propose a novel cross-organisms learning approach to reliably predict new functionalities for the genes of an organism based on the known controlled annotations of the genes of another, evolutionarily related and better studied, organism. We leverage a new representation of the annotation discovery problem and a random perturbation of the available controlled annotations to allow the application of supervised algorithms to predict with good accuracy unknown gene annotations. Taking advantage of the numerous gene annotations available for a well-studied organism, our cross-organisms learning method creates and trains better prediction models, which can then be applied to predict new gene annotations of a target organism. We tested and compared our method with the equivalent single organism approach on different gene annotation datasets of five evolutionarily related organisms (Homo sapiens, Mus musculus, Bos taurus, Gallus gallus and Dictyostelium discoideum). Results show both the usefulness of the perturbation method of available annotations for better prediction model training and a great improvement of the cross-organism models with respect to the single-organism ones, without influence of the evolutionary distance between the considered organisms. The generated ranked lists of reliably predicted annotations, which describe novel gene functionalities and have an associated likelihood value, are very valuable both to complement available annotations, for better coverage in biomedical knowledge discovery analyses, and to quicken the annotation curation process, by focusing it on the prioritized novel annotations predicted.


Catechol-bearing imidazolium and benzimidazolium chlorides as promising antimicrobial agents.
Catechol-containing imidazolium (four) and benzimidazolium chlorides (eight) were synthesized to evaluate their antimicrobial properties. All the compounds were fully characterized using 1 H and 13 C nuclear magnetic resonance, liquid chromatography-mass spectrometry, infrared spectroscopic methods, and elemental analyses. Antimicrobial activities of the compounds were tested against the bacteria Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Enterococcus faecalis, and the fungal strains Candida albicans and Candida glabrata, and promising results were achieved. The two most important benzyl-substituted benzimidazolium chlorides, 3l and 3k, showed comparable activity to vancomycin against MRSA.