A receptor-binding-based bioassay to determine the potency of a plasmid biopharmaceutical encoding VEGF-C.
Over the last decade biological assays (bioassays) have gained much importance for quality control in biopharmaceutical development and manufacturing. Here we describe the development and validation of a bioassay to determine the biological activity (potency) of the plasmid biopharmaceutical pVGI.1 which encodes the VEGF-C (VEGF-2) protein. This assay was developed to test drug substance and drug product for release and stability testing for phase I and II clinical trials. The main focus was on fast assay development and easy handling of the assay, combined with valid results representing the specific therapeutic mechanism. The method includes the expression of the VEGF-C protein in mammalian cells and its binding to the cell surface receptor VEGFR-3. The binding activity of VEGF-C to its immobilized receptor is quantified in a colorimetric assay. IC50 values of VEGF-C expressed after transfection with sample plasmid and an in-house standard plasmid are determined. The ratio of the IC50 value of the test article to that of the reference standard reflects the potency of the sample. The potency assay meets the criteria generally requested by authorities for precision, linearity, accuracy, and range. Therefore the assay can be used in pharmaceutical quality control and is a suitable basis for development of related bioassays.


Hemorrhagic shock successfully treated with emergency pancreaticoduodenectomy for pancreatic carcinoma in radiochemotherapy: A case report.
Pancreatic cancer often invades the duodenum; however, it rarely causes duodenal bleeding. We describe a case of a 77-year-old Japanese woman admitted to our hospital with hematemesis, who presented with pancreatic head cancer and received radiochemotherapy (radiotherapy + gemcitabine). The following day, she developed hemorrhagic shock, and an emergency endoscopy was performed, which revealed a bleeding ulcerative lesion in the second portion of the duodenum. We chose surgical treatment over other therapies (interventional radiology or endoscopy). Pancreaticoduodenectomy was successfully performed to control hemorrhage and the Child's method was used for reconstruction. The patient's postoperative course was uneventful. After her condition improved, she was treated for residual cancer 2 months after surgical treatment; therefore, complementary radiation with concurrent chemotherapy based on GEM was administrated. However, she died 12 months after the surgery. During the treatment of pancreatic cancer, it is necessary to avoid bleeding as much as possible by considering prophylactic treatment, including periodic gastrointestinal scrutiny and resection or embolization, depending on the case.


Ventilation with biphasic positive airway pressure in experimental lung injury. Influence of transpulmonary pressure on gas exchange and haemodynamics.
We investigated whether improvement in ventilation perfusion (.V(A)/.Q) distribution during mechanical ventilation using biphasic positive airway pressure (BIPAP) with spontaneous breathing may be attributed to an effectively increased transpulmonary pressure (P(TP)) and can also be achieved by increasing P(TP) during controlled ventilation. In 12 pigs with saline lavage-induced lung injury we compared the effects of BIPAP to pressure-controlled ventilation with equal airway pressure (PCV(AW)) or equal transpulmonary pressure (PCV(TP)) on V(A)/.Q distribution assessed by the multiple inert gas elimination technique (MIGET). Animal laboratory study. Intrapulmonary shunt was 33+/-11% during BIPAP, 36+/-10% during PCV(AW) and 33+/-15% during PCV(TP) ( p= n.s.). BIPAP resulted in higher PaO(2) than PCV(AW) (188+/-83 versus 147+/-82 mmHg, p < 0.05), but not than PCV(TP) (187+/-139 mmHg). Oxygen delivery was significantly higher during BIPAP (530+/-109 ml/min) versus 374+/-113 ml/min during PCV(AW) and 353+/-93 ml/min during PCV(TP) ( p < 0.005). Tidal volume with PCV(TP) increased to 11.9+/-2.3 ml/kg, compared to 8.5+/-0.8 with BIPAP and 7.6+/-1.4 with PCV(AW) ( p <0.001) and cardiac output decreased to 3.5+/-0.6 l/min (BIPAP 4.9+/-0.8 and PCV(AW) 3.9+/-0.8, p<0.006). In experimental lung injury, BIPAP with preserved spontaneous breathing was effective in increasing regional P(TP), since pressure-controlled ventilation with the same P(TP) resulted in similar gas exchange effects. However, PCV(TP) caused increased airway pressures and tidal volumes, whereby, with BIPAP, less depression of oxygen delivery and cardiac output were observed. BIPAP could be useful in maintaining pulmonary gas exchange and slightly improving oxygenation without interfering with circulation as strongly as PCV does.


Evaluation of a urinary catheter with a preconnected closed drainage bag.
The incidence of bacteriuria, significant urinary tract infection, and cost associated with the use of two urinary catheter drainage systems were evaluated in a population of hospitalized adult males. A commercially available device comprised of a Foley catheter attached with a heat-shrunk plastic seal to the tubing of a closed drainage bag (preconnected system) was compared with a standard system that had the Foley catheter attached to the closed drainage bag after it had been inserted. Using a randomized prospective design, the performance of the preconnected system in 97 patients was compared with that of the standard system in 105 patients. Catheters were left in place a mean of 6.4 and 7.6 days in the respective groups. Bacteriuria occurred after catheter insertion in 11.3% of patients receiving the preconnected system and 13.3% of patients receiving the standard system (not statistically significant). When bacteriuria developed, it occurred within seven days of catheter insertion in 50% of instances, irrespective of drainage system employed, suggesting that manipulations related to catheter insertion were important in initiating bacteriuria. Significant urinary tract infections occurred in only 2% of all patients studied. The higher cost for purchasing the preconnected system was not warranted for the population of patients studied.


Melittin-induced alterations in dynamic properties of human red blood cell membranes.
The interaction of bee venom melittin with erythrocyte membrane ghosts has been investigated by means of fluorescence quenching of membrane tryptophan residues, fluorescence polarization and ESR spectroscopy. It has been revealed that melittin induces the disorders in lipid-protein matrix both in the hydrophobic core of bilayer and at the polar/non-polar interface of melittin complexed with erythrocyte membranes. The peptide has been found to act most efficiently at the concentration of the order of 10(-10) mol/mg membrane protein. The apparent distance separating the membrane tryptophan and bound 1-anilino-8-naphthalenesulphonate (ANS) molecules is decreased upon melittin binding, which results in a significant increase of the maximum energy transfer efficiency. Significant changes in the fluorescence anisotropy of both 1,6-diphenyl-1,3,5-hexatriene and 1-anilino-8-naphthalenesulphonate bound to erythrocyte ghosts, which have been observed in the presence of melittin and crude venom, indicate membrane lipid bilayer rigidization. The effect of crude honey bee venom has been found to be of similar magnitude as the effect of pure melittin at the concentration of 10(-10) mol/mg membrane protein. Using two lipophilic spin labels, methyl 5-doxylpalmitate and 16-doxylstearic acid, we found that melittin at its increasing concentrations induces a well marked rigidization in the deeper regions of lipid bilayer, whereas the effect of rigidization near the membrane surface maximizes at the melittin concentration of 10(-10) mol/mg (10(-4) mol melittin per mole of membrane phospholipid). The decrease in the ratio hw/hs of maleimide and the rise in relative rotational correlation time (tau c) of iodacetamid spin label, indicate that melittin effectively immobilizes membrane proteins in the plane of the lipid bilayer. We conclude that melittin-induced rigidization of the lipid bilayer may induce a reorganization of lipid assemblies as well as the rearrangements in membrane protein pattern and consequently the alterations in lipid-protein interactions. Thus, the interaction of melittin with erythrocyte membranes is supposed to produce local conformational changes in membranes, which are discussed in the connection with their significance during the synergistic action of melittin and phospholipase of bee venom on red blood cells.


Amniotic membrane as a carrier for cultivated human corneal endothelial cell transplantation.
It would be advantageous if cultivated human corneal endothelial cells (cHCECs) could be transplanted for the treatment of diseases caused by corneal endothelial disorders. To achieve this, a matrix that can serve as a carrier for cHCECs is needed. The present study was conducted to examine the feasibility of using amniotic membrane (AM) as a carrier for this application. HCECs obtained from peripheral corneal tissue were cultivated, passaged, and transplanted onto denuded AM. The cell density and morphology of the resultant cHCECs on AM were examined by light, scanning electron, and transmission electron microscopy. To determine whether these cHCEC sheets on AM carrier were functional in vivo, the cHCEC sheets on AM were transplanted onto rabbit corneas whose Descemet's membrane and endothelial cells had been completely removed. After transplantation, the corneal appearance was examined by slit lamp biomicroscopy, and corneal thickness was measured daily by pachymetry. At 7 days after surgery, the grafts were examined by light, scanning electron, and transmission electron microscopy. The density of the cHCECs on AM was greater than 3000 cells/mm(2). Morphologically, the cHCEC sheets consisted of a fairly continuous layer of flat squamous polygonal endothelial cells that appeared uniform in size with tightly opposed cell junctions in vitro and in vivo after transplantation. The corneas that received transplanted cHCEC sheets had little edema and retained their thinness and transparency. The cell density and morphology of cHCECs on AM were similar to those of normal corneas, and cHCECs on AM were functional in vivo. These results indicate that AM maintains HCEC morphology and function and could serve as a carrier for cHCEC transplantation.


Metabolism of 1-alkyl-2-acyl-GPC in human platelets in response to stimulation by thrombin.
Washed human platelets were incubated with radioactive 1-[3H]alkyl-2-hydroxyglycero-3-phosphocholine (lyso-PAF) at 37 degrees C. [3H]lyso-PAF was converted by platelets into [3H]alkylacyl-GPC which was incorporated. Incorporation of radioactivity was time dependent and reached a maximum of 57 percent in one h. This formation and incorporation of [3H]alkylacyl-GPC was inhibited (50%) by extracellular calcium (1.3 mM). Labeled platelets were treated for 5 min with either thrombin (2.5 U/ml) or saline solution. While there was no change in the saline control, thrombin induced a reduction in the content of [3H]alkylacyl-GPC, accompanied by an increase in [3H]lyso-PAF presumably by stimulation of phospholipase A2. There was no apparent increase in radioactivity comigrating with PAF. This was probably due to the overwhelming dilution of the radioactive alkylacyl-GPC by the endogenous nonradioactive compound (ratio-1/3200). These studies suggest that human platelets can take up lyso-PAF and acylate it to alkylacyl-GPC which is susceptible to phospholipase A2 activity.


Association between the cholesteryl ester transfer protein gene and polypoidal choroidal vasculopathy.
To determine whether genetic variants in the lipid-associated genes are related to the risk of developing polypoidal choroidal vasculopathy (PCV) in a Japanese population. Five hundred eighty-one patients with PCV and 793 controls were enrolled in the study. Association analysis of allele and genotype frequencies was performed for the following single-nucleotide polymorphisms (SNPs) that are associated with high-density lipoprotein cholesterol levels in blood: rs493258 at the hepatic lipase gene (LIPC), rs3764261 at the cholesteryl ester transfer protein gene (CETP), and rs12678919 at the lipoprotein lipase gene (LPL). A further model adjusting for age-related maculopathy susceptibility 2 (ARMS2) A69S, complement factor H (CFH) I62V, age, sex, and smoking status was used to confirm the independent association of these SNPs with other covariates. CETP rs3764261 was significantly associated with the development of PCV; the frequency of the minor allele A was higher in the PCV cases (24.0%) than in the control subjects (18.5%) (P = 0.0025; odds ratio [OR], 1.41; 95% confidence interval, 1.13-1.75). Furthermore, we found an independent association of CETP variants with age, sex, smoking status, and genetic background of ARMS2 A69S, CFH I62V, LIPC rs493258, and LPL rs12678919 (P = 0.0013; OR, 1.50). LIPC rs493258 and LPL rs12678919 did not show significant associations with the development of PCV (P > 0.05). CETP variants are associated a risk of developing PCV among the Japanese population.


The implementation of best practice in medication administration across a health network: a multisite evidence-based audit and feedback project.
Medication errors present a significant risk to patient safety. The "rights" of medication administration represent one approach to potentially reducing this risk. The aim of this project was to implement an evidence-based audit and feedback project to improve compliance with best practice in this area across a health network. A baseline audit was conducted to determine compliance with evidence-based standards by trained observers. The results of this audit were analysed and fed back to staff. An analysis of barriers to compliance was undertaken by key staff within the organization, which was followed by the implementation of targeted strategies to improve compliance. A follow-up audit was conducted and the results compared to the baseline audit. There were improvements in the percentage of compliance across all of the eight criteria audited, with statistically significant improvements found in six of the eight. In general, compliance with the criteria was high in both the baseline and follow-up audits. This audit and feedback implementation project was successful in increasing compliance and knowledge in this area and providing future direction for sustaining evidence-based practice change. It is now planned to use this approach for rolling out future implementation projects within this health system.


Maternal and newborn healthcare utilization in Kampala urban slums: perspectives of women, their spouses, and healthcare providers.
It is assumed that the health conditions of urban women are superior to their rural counterparts. However, evidence from Asia and Africa, show that poor urban women and their families have worse access to antenatal care and facility childbirth compared to the rural women. The maternal, newborn, and child mortality rates as high as or higher than those in rural areas. In Uganda, maternal and newborn health data reflect similar trend. The aim of the study was to understand factors that influence use of maternal and newborn healthcare in two urban slums of Kampala, Uganda. A qualitative study was conducted in urban slums of Kampala, Uganda and conducted 60 in-depth interviews with women who had given birth in the 12 months prior to data collection and traditional birth attendants, 23 key informant interviews with healthcare providers, coordinator of emergency ambulances/emergency medical technicians and the Kampala Capital City Authority health team, and 15 focus group discussions with partners of women who gave birth 12 months prior to data collection and community leaders. Data were thematically coded and analyzed using NVivo version 10 software. The main determinants that influenced access to and use of maternal and newborn health care in the slum communities included knowledge about when to seek care, decision-making power, financial ability, prior experience with the healthcare system, and the quality of care provided. Private facilities were perceived to be of higher quality, however women primarily sought care at public health facilities due to financial constraints. Reports of disrespectful treatment, neglect, and financial bribes by providers were common and linked to negative childbirth experiences. The lack of adequate infrastructure and basic medical equipment and medicine impacted patient experiences and provider ability to deliver quality care. Despite availability of healthcare, urban women and their families are burdened by the financial costs of health care. Disrespectful and abusive treatment at hands of healthcare providers is common translating to negative healthcare experiences for women. There is a need to invest in quality of care through financial assistance programs, infrastructure improvements, and higher standards of provider accountability are needed.


Dual roles of the 90-kDa heat shock protein hsp90 in modulating functional activities of the dioxin receptor. Evidence that the dioxin receptor functionally belongs to a subclass of nuclear receptors which require hsp90 both for ligand binding activity and repression of intrinsic DNA binding activity.
Signal transduction by dioxin (2,3,7,8-tetrachloro-dibenzo-p-dioxin) is mediated by the intracellular dioxin receptor which, in its dioxin-activated state, regulates transcription of target genes encoding drug metabolizing enzymes such as cytochrome P-450IA1 and glutathione S-transferase Ya. Upon binding of dioxin the receptor translocates from the cytoplasm to the nucleus in vivo and is converted from a latent non-DNA binding form to a species which binds to dioxin-responsive positive control elements in vitro. The latent receptor form is associated with an inhibitory protein (the 90-kDa heat shock protein, hsp90), the release of which is necessary to unmask the DNA binding activity of the receptor. Here we have established a protocol to disrupt the hsp90-receptor complex in the absence of ligand. We show that it was possible to covalently cross-link with dioxin only the hsp90-associated form of dioxin receptor. In contrast, the disrupted hsp90-free form of receptor did not form a stable complex with dioxin but bound DNA constitutively. Moreover, we could partially reconstitute the ligand binding activity of the salt-disrupted hsp90-free dioxin receptor by incubation with hsp90-containing reticulocyte lysate but not by incubation with wheat germ lysate which lacks immuno-detectable levels of hsp90. Thus, we demonstrate that the dioxin receptor loses its high affinity ligand binding activity following release of hsp90 and that it is possible to reverse this process. In conclusion, hsp90 appears to play dual roles in the modulation of functional activities of the dioxin receptor: (i) it represses the intrinsic DNA binding activity of the receptor and (ii) it appears to determine the ability of the receptor to assume and/or maintain a ligand binding conformation.


Accurate SAXS profile computation and its assessment by contrast variation experiments.
A major challenge in structural biology is to characterize structures of proteins and their assemblies in solution. At low resolution, such a characterization may be achieved by small angle x-ray scattering (SAXS). Because SAXS analyses often require comparing profiles calculated from many atomic models against those determined by experiment, rapid and accurate profile computation from molecular structures is needed. We developed fast open-source x-ray scattering (FoXS) for profile computation. To match the experimental profile within the experimental noise, FoXS explicitly computes all interatomic distances and implicitly models the first hydration layer of the molecule. For assessing the accuracy of the modeled hydration layer, we performed contrast variation experiments for glucose isomerase and lysozyme, and found that FoXS can accurately represent density changes of this layer. The hydration layer model was also compared with a SAXS profile calculated for the explicit water molecules in the high-resolution structures of glucose isomerase and lysozyme. We tested FoXS on eleven protein, one DNA, and two RNA structures, revealing superior accuracy and speed versus CRYSOL, AquaSAXS, the Zernike polynomials-based method, and Fast-SAXS-pro. In addition, we demonstrated a significant correlation of the SAXS score with the accuracy of a structural model. Moreover, FoXS utility for analyzing heterogeneous samples was demonstrated for intrinsically flexible XLF-XRCC4 filaments and Ligase III-DNA complex. FoXS is extensively used as a standalone web server as a component of integrative structure determination by programs IMP, Chimera, and BILBOMD, as well as in other applications that require rapidly and accurately calculated SAXS profiles.


The effect of famotidine, a new H2 receptor antagonist, on antipyrine clearance in healthy volunteers.
The salivary clearance of Antipyrine (15 mg/kg p.o.) has been measured in nine healthy volunteers (5 males, 4 females) before and after 7 days treatment with Famotidine (40 mg/day at bedtime). Samples of saliva were collected twenty four hours after each dose of Antipyrine. The post-treatment sample was taken 12 hours after the last dose of Famotidine. The clearance of Antipyrine was measured by Gas-Liquid Chromatography using the simplified formula proposed by Dossing et al. (1982). After treatment with Famotidine, a slight but significant (p less than 0.01) increase of Antipyrine clearance was observed in eight out of the nine subjects, suggesting some interference of Famotidine with Antipyrine metabolism.


Voltage oscillations in mammalian metaphase II oocytes.
The membrane potential has been measured in ovulated mouse oocytes using conventional electrophysiological techniques. Temporal oscillations in membrane voltage have been observed in the oocytes, with periods of about 6 h. This oscillatory pattern, peculiar to oocytes in metaphase II, might explain the differences in membrane potential values reported in several studies on mammalian oocytes.


Efficacy of chlorine dioxide disinfection to non-fermentative Gram-negative bacilli and non-tuberculous mycobacteria in a hospital water system.
Chlorinated tap water in hospitals often contains low levels of non-fermentative Gram-negative bacilli (NFGNB) and non-tuberculous mycobacteria (NTM). Measures are needed to ensure a safe water supply in hospitals to prevent nosocomial infections from these waterborne pathogens. To evaluate the efficacy of ClO2 treatment of a hospital water system on the levels of NFGNB and NTM in the water. Our institution is a 1000-bed medical centre with two main buildings (B1 and B2). B1 has three intensive care units (ICUs) and transplant wards and polyethylene water pipes. B2 (control) has no ICUs and galvanized water pipes. A ClO2 generating unit was installed in the water system of B1 in April 2012 and water samples were collected in B1 and B2 before and eight times after installation. All samples were cultured for NFGNB and NTM. The ClO2 concentration was significantly lower in the hot water than in the cold water (P<0.001). After 40 weeks of ClO2 use, the overall NFGNB colonies decreased significantly (hot water: 160±143 vs 2±4cfu/mL, P<0.001; cold water: 108±138 vs 3±7cfu/mL, P<0.001). Highly prevalent nosocomial NFGNB, such as Pseudomonas spp. and Stenotrophomonas spp., were undetected three months after ClO2 disinfection; Sphingomonas spp. persisted but had lower colony counts. NTM was present in 25% (three out of 12) of sampling locations initially, but was not detected at two weeks after ClO2 disinfection. The ICUs had no overall change in the number of NFGNB nosocomial infections after the intervention. Addition of a ClO2 disinfection unit to our hospital water system reduced the numbers of NTM and NFGNB in the hot and cold water systems.


Huangqin Qingre Qubi Capsule inhibits RA pathology by binding FZD8 and further inhibiting the activity of Wnt/β-catenin signaling pathway.
Huangqin Qingre Qubi Capsule (HQC) is a Chinese herbal compound for the treatment of rheumatoid arthritis (RA), which is made from dry roots of Scutellaria baicalensis Georgi, dry mature seeds of Gardenia jasminoides J.Ellis, dry and mature seeds of Coix lacryma-jobi var. stenocarpa Oliv., dry mature seeds of Amygdalus persica L. and roots and rhizomes of Clematis chinensis Osbeck in the proportion of 10:9:30:5:10. HQC has a significant effect in clinical treatment of RA, which can inhibit RA inflammation, improve oxidative stress state, and effectively relieve symptoms of RA patients. The anti-arthritis effect of HQC and its mechanism, especially whether it improves RA through FZD8-Wnt/β-catenin signal axis, were studied using adjuvant arthritis (AA) rats and FLS from RA patients. Real time qPCR (RT-qPCR), Western blot (WB), confocal microscopy and other molecular biological methods were used to study the anti-RA effect of HQC and its mechanism. The expression of FZD8 was significantly up-regulated in synovium and FLS of AA rats and RA FLS. FZD8 significantly activated the Wnt/β-catenin signaling pathway, promoted abnormal proliferation of FLS, increased the levels of inflammatory factors IL-1β, IL-6 and IL-8, and significantly increased the expression of matrix metalloproteinase 3 (MMP3) and fibronectin. HQC has significant therapeutic effect on AA rats. Molecular docking and molecular dynamics showed that HQC had a good binding ability with FZD8. We also confirmed that HQC inhibited Wnt/β-catenin signaling pathway by binding FZD8, and reduced the levels of the above inflammatory factors and pathological genes of RA. The expression of FZD8 is significantly increased in AA rats and FLS from RA patients. Clarify that HQC improves RA through the FZD8-Wnt/β-catenin signal axis, provide a clear therapeutic mechanism for HQC to improve RA, and also provide a basis for clinical promotion of HQC.


B,B-Diporphyrinbenzyloxy-BODIPY dyes: synthesis and antenna effect.
B,B-Diporphyrinbenzyloxy-BODIPY derivatives have been prepared in high yields, and the photophysical properties are reported. Singlet energy transfers from BODIPY to the porphyrin units have been analyzed.


Design and development of polymers for gene delivery.
The lack of safe and efficient gene-delivery methods is a limiting obstacle to human gene therapy. Synthetic gene-delivery agents, although safer than recombinant viruses, generally do not possess the required efficacy. In recent years, a variety of effective polymers have been designed specifically for gene delivery, and much has been learned about their structure-function relationships. With the growing understanding of polymer gene-delivery mechanisms and continued efforts of creative polymer chemists, it is likely that polymer-based gene-delivery systems will become an important tool for human gene therapy.


Mechanical deterioration underlies malignant behavior of aneurysmal human ascending aorta.
The human ascending aorta becomes markedly prone to rupture and dissection at a diameter of 6 cm. The mechanical substrate for this malignant behavior is unknown. This investigation applied engineering analysis to human ascending aortic aneurysms and compared their structural characteristics with those of normal aortas. We measured the mechanical characteristics of the aorta by direct epiaortic echocardiography at the time of surgery in 33 patients with ascending aortic aneurysm undergoing aortic replacement and in 20 control patients with normal aortas undergoing coronary artery bypass grafting. Six parameters were measured in all patients: aortic diameter in systole and diastole, aortic wall thickness in systole and diastole, and blood pressure in systole and diastole. These were used to calculate mechanical characteristics of the aorta from standard equations. Aortic distensibility reflects the elastic qualities of the aorta. Aortic wall stress reflects the disrupting force experienced within the aortic wall. Incremental elastic modulus indicates loss of elasticity reserve. Aortic distensibility falls to extremely low levels as aortic dimension rises toward 6 cm (3.02 mm Hg(-1) for small aortas versus 1.45 mm Hg(-1) for aortas larger than 5 cm, P < .05). Aortic wall stress rises to 157.8 kPa for the aneurysmal aorta, compared with 92.5 kPa for normal aortas. For 6-cm aortas at pressures of 200 mm Hg or more, wall stress rises to 857 kPa, nearly exceeding the known maximal tensile strength of human aneurysmal aortic wall. Incremental elastic modulus deteriorates (1.93 +/- 0.88 MPa vs 1.18 +/- 0.21 MPa, P < .05) in aneurysmal aortas relative to that in normal aortas. The mechanical properties of the aneurysmal aorta deteriorate dramatically as the aorta enlarges, reaching critical levels associated with rupture by a diameter of 6 cm. This mechanical deterioration provides an explanation in engineering terms for the malignant clinical behavior (rupture and dissection) of the aorta at these dimensions. This work adds to our fundamental understanding of the biology of aortic aneurysms and promises to permit future application of engineering measurements to supplement aneurysm size in clinical decision making in aneurysmal disease.


The phenotype of polycystic ovary syndrome ameliorates with aging.
To assess the effects of aging on the features of polycystic ovary syndrome (PCOS). Retrospective longitudinal follow-up study. Tertiary care center. Patients with PCOS, diagnosed according to the 2003 Rotterdam criteria, who visited the outpatient clinic on consecutive occasions with a minimum interval of 6 months. Comparisons were made between the first visit and the consecutive visit grouped by intervals. Changes in clinical and endocrine characteristics. A total of 254 women visited the outpatient clinic on 2 occasions each. Consecutive visits were grouped into 0.5 to 3.9 years (n = 172; mean follow-up, 2.6 years) and 4.0 to 7.0 years (n = 82; mean follow-up, 5.5 years). At their second visit, significantly more women had regained a regular cycle. The total antral follicle count was similar. Serum levels of testosterone, androstenedione, and dehydroepiandrosterone sulfate had decreased significantly. Plasma glucose levels had increased, whereas serum insulin levels and homeostasis model assessment score had significantly decreased. The PCOS phenotype changed with aging, suggesting an amelioration of the phenotype and ovarian dysfunction as indicated by the increase in number of regular menstrual cycles, decrease in serum androgen levels, and decrease in insulin resistance.


Fine structure of the dorsal cap of the inferior olive and its GABAergic and non-GABAergic input from the nucleus prepositus hypoglossi in rat and rabbit.
The dorsal cap of the inferior olive is involved in the control of eye movements and is excited by inputs from the midbrain. In the present study we attempted to determine the inhibitory input to this nucleus in rat and rabbit. The projection from the nucleus prepositus hypoglossi to the dorsal cap was studied in the light microscope by anterograde tracing of Phaseolus vulgaris-leucoagglutinin and lesion-induced depletion of glutamic acid decarboxylase immunoreactivity, and in the electron microscope by anterograde tracing of wheat germ agglutinin-coupled horseradish peroxidase combined with GABA immunocytochemistry. We show that the nucleus prepositus hypoglossi projects bilaterally to the dorsal cap, contralaterally to the ventrolateral outgrowth, and ipsilaterally to the medial accessory olive. After lesioning of the nucleus prepositus hypoglossi, the caudal dorsal cap was depleted of most of its glutamic acid decarboxylase-immunoreactive terminals while the rostral dorsal cap and the ventrolateral outgrowth were depleted of a minor part. Ultrastructural analysis indicates that the majority, but not all, of the terminals from the nucleus prepositus hypoglossi in the dorsal cap are GABA-positive. These GABA-positive and GABA-negative terminals form predominantly symmetric and asymmetric synapses; most of them synapse on dendrites outside and inside glomeruli, frequently in association with dendrodendritic gap junctions, while a small minority are axosomatic. None of the terminals from the nucleus prepositus hypoglossi was found to form a crest synapse, although synapses of this kind were predominantly formed by GABAergic terminals. This study shows that the dorsal cap receives a major inhibitory input from the nucleus prepositus hypoglossi, the terminals of which are located at strategic positions on the olivary neurons.


Office screening for asymptomatic urinary tract infections. The evaluation of a practical, economic and reliable method screening the urines of asymptomatic girls in a busy pediatric office.
The urines of 3,270 asymptomatic girls were screened at annual physical examinations. The urines were collected in Dixie Cups without prior preparation of the perineum and cultured on 5% sheep cell agar. Less than 2 per cent of the specimens showed significant growth, thus requiring no follow-up visits by 98 per cent of the patients. Slightly less that 1/2 of one per cent were found to have asymptomatic urinary tract infections. The procedure was found to be practical, economically feasible and reliable, and was well accepted by both the parents and patients.


Effects of selenium intake on the expression of prostaglandin-endoperoxide synthase 2 (cyclooxygenase-2) and matrix metallopeptidase-9 genes in the coronary artery disease: Selenegene study, a double-blind randomized controlled trial.
The oxidative stress is regarded as one of the main contributors to the health problem. Cyclooxygenase-2 (COX-2) and matrix metallopeptidase-9 (MMP-9) are two of the important genes that are reported to be involved in the cardiovascular disease (CVD) development in the molecular and genetic association studies. The aim of this study was to evaluate the level of expression of COX-2 and MMP-9 after selenium supplementation in patients with coronary artery disease (CAD). In this sub-study of Selenegene study, subjects were randomly divided into groups, 19 subjects who received selenium and 22 patients with CAD who received placebo. Patients received either 200-mg selenium yeast tablets or placebo tablets after a meal, once daily for 60 days. The messenger ribonucleic acid (mRNA) levels of the selenium and prostaglandin-endoperoxide synthase 2 (PTGS2) (COX-2) and MMP-9 genes products were determined before and after the study. In this sub-study, 41 Iranian patients with CVD were enrolled (placebo group: n = 22, selenium intervention: n = 19). Fasting blood sugar (FBS) was higher among placebo group than selenium group (93.4 ± 12.7 vs. 124.4 ± 40.6 mg/dl, P = 0.03). Triglyceride (TG) level was higher among selenium group versus placebo group (123.3 ± 34.0 vs. 184.8 ± 69.4 mg/dl, P = 0.006). The data analysis demonstrated that the expression of MMP-9 and COX-2 genes did not change significantly in both selenium and placebo groups. This study showed a positive association between the expression of MMP-9 and COX-2 in the patients with CAD who received selenium but not the placebo groups. Yet, these findings need to be confirmed in further details and expanded sample size.


Impact of a multidimensional approach on ventilator-associated pneumonia rates in a hospital of Shanghai: findings of the International Nosocomial Infection Control Consortium.
The aim of this study was to analyze the impact of a multidimensional infection control approach on the reduction of ventilator-associated pneumonia (VAP) in intensive care units (ICUs) patients of one hospital in China. We conducted a before-after study from January 2005 to July 2009, which was divided into baseline (phase 1) and intervention (phase 2) periods. During phase 1, active prospective outcome surveillance of VAP was performed by applying the definitions of the Centers for Disease Control and Prevention/National Health Safety Network, and the methodology of the International Nosocomial Infection Control Consortium. During phase 2, the multidimensional approach was implemented. Ventilator-associated pneumonia rates obtained in phases 1 and 2 were compared in yearly periods. We recorded data from 16,429 patients hospitalized in 3 ICUs, for a total of 74,116 ICU bed days. The VAP baseline rate was 24.1 per 1000 ventilator-days. During phase 2, the VAP rate significantly decreased to 5.7 per 1000 ventilator-days in 2009 (2009 vs 2005: relative risk, 0.31; 95% confidence interval, 0.16-0.36; P = .0001), amounting to a 79% cumulative VAP rate reduction. Implementing a multidimensional infection control intervention for VAP was associated with a significant cumulative reduction in the VAP rate in our ICUs.


Determinants of condom uptake among HIV voluntary counselling and testing clients: experiences from a hospital-based study in south India.
HIV voluntary counselling and testing was a key HIV prevention strategy brought to scale by India's National AIDS Control Organization. Condom uptake is an essential metric of intervention impact given the expansion of the epidemic into an increasingly diverse population. With only 20% of first-time counselling and testing clients at the largest HIV treatment hospital in south India reporting previous condom use, the question of intervention impact on condom use deserves investigation. In this study, we track intervention impact across various demographic groups and identify the added value of more thorough counselling. Data were collected from 8,865 individuals who attended counselling multiple times at the Tamil Nadu Government Hospital of Thoracic Medicine over the years 2004-2009. Counsellors recorded client demographic characteristics, HIV risk behaviours reported, and counselling services provided after each counselling session. Matching and regression methods were used to determine the probability of condom uptake by serostatus, gender, and receipt of personalized risk reduction counselling while controlling for other characteristics. HIV counselling and testing was associated with condom uptake among 29.2% of HIV positive women (CI 24.5-34.4%), 31.7% of HIV positive men (CI 27.8-35.4%), 15.5% of HIV negative women (CI 11.2-20.8%), and only 3.6% of HIV negative men (CI 1.9-5.9%) who had previously never used condoms. Personalized risk reduction counselling increased impact in some groups; for example an additional 18% of HIV negative women (CI 11.3-24.4%) and 17% of HIV positive men (CI 10.9-23.4%) started using condoms. The number of sexual partners was not associated with the impact of counselling completeness. Because the components of testing and counselling impact the condom use habits of men and women differently, understanding the dynamics of condom use negotiation between partners is essential to optimizing impact on Indian couples. Clients' predicted condom uptake ranged between 4% and 47% depending on factors like gender, serostatus, and services provided. Personalized risk reduction counselling is associated with increased chance of condom use, with larger gains in HIV negative women and HIV positive men. HIV negative men are least likely to start using condoms and least impacted by additional counselling.


Distinct domains of Bcl-XL are involved in Bax and Bad antagonism and in apoptosis inhibition.
Pro-survival factor Bcl-X(L) can antagonize the pro-apoptotic functions of Bax and Bad via two distinct mechanisms. It can block Bax-mediated cell death by preventing Bax translocation from the cytosol to mitochondria. On the other hand, Bcl-X(L) can neutralize Bad by sequestering it to mitochondria. In order to map the domains of Bcl-X(L) involved in inhibiting Bax and Bad, we have carried out mutational analyses of this protein. This was done by deleting the key domains of Bcl-X(L), including its BH1-4 domains, the flexible loop, the C-terminal hydrophobic domain, and segments of the alpha5-alpha6 hairpin. The resulting Bcl-X(L) mutant constructs were then co-transfected with either GFP-Bax or GFP-Bad. We found that the BH1-4 domains and the C-terminal segment of Bcl-X(L) were essential for blocking Bax localization to mitochondria. On the other hand, only its BH1 and BH3 domains and the C-terminal hydrophobic segment were necessary for sequestering Bad to mitochondria. In addition, by immunoprecipitation analyses, we found that these deletions differentially affected the ability of the Bcl-X(L) mutant proteins to bind Bax and Bad. Finally, cell viability assays indicated that the BH1-4 domains of Bcl-X(L) were the primary domains required for inhibiting staurosporine-induced apoptosis, suggesting that distinct domains of Bcl-X(L) are involved in antagonizing Bax and Bad and in apoptosis inhibition.


Effects of dietary butyrate supplementation and oral nonsteroidal antiinflammatory drug administration on uterine inflammation and interval to first ovulation in postpartum dairy cows.
This study evaluated the effects of dietary butyrate supplementation and oral nonsteroidal antiinflammatory drug (NSAID) administration on uterine inflammation and the interval from calving to first ovulation (ICFO; in days). We hypothesized that a combination of dietary butyrate and oral NSAID would reduce uterine inflammation and decrease ICFO. Sixty-five cows were enrolled in a 2 × 2 factorial design and assigned to receive an iso-energetic diet containing a supplement of either butyrate (fatty acid-coated calcium butyrate) or control (commercial fat and calcium carbonate mixture) at 1.42% of diet dry matter, during the calving transition period from -28 (±3) to +24 (±3) days in milk (DIM; calving = d 0). At 12 to 24 h postcalving, cows received an oral NSAID (1 mg of meloxicam/kg of BW) or a placebo (food dye). Ovarian ultrasonography was performed weekly from 14 DIM until first ovulation or up to 56 DIM. Endometrial cytology was performed at 28 DIM to assess uterine inflammation based on polymorphonuclear leukocytes (PMN). No interactions were detected between treatments. The proportions of cows with high (>18%) endometrial PMN did not differ between butyrate and control diets or between NSAID and placebo. Mean (± standard error of mean) ICFO did not differ between butyrate (28 ± 2 d) and control (25 ± 2 d) or between NSAID (29 ± 2 d) and placebo (24 ± 2 d). However, the ovulation rate up to 56 DIM (hazard ratio: 0.61; 95% confidence interval: 0.35 to 1.04) established by survival analysis tended to be lower in NSAID than in placebo. In conclusion, dietary butyrate supplementation and oral NSAID administration did not reduce endometrial inflammation or the mean ICFO, but NSAID-treated cows tended to have a lower rate of ovulation up to 56 DIM.


About the large fluctuations observed using gas-phase molecular dynamics in the K-ras gene containing a mismatch.
We describe how we can reduce the periodic bending motions in the simulation in vacuo of the molecular dynamics of a short DNA fragment containing the Gly 12 hot spot of the K-ras oncogene and having at its center a mismatch CA+.


Ecology of SO2 resistance : IV. Predicting metabolic responses of fumigated shrubs and trees.
10 broadleafed trees and shrubs native to the mediterranean climactic zone in California were surveyed for their photosynthetic and stomatal responses to SO2. These species ranged from drought deciduous to evergreen and had diverse responses to SO2. These results suggest an approach for predicting SO2 resistances of plants.We found that conductance values of plants in SO2-free air can be used to estimate the quantity of SO2 which plants absorb. These estimates are based on conductance values for plants in non-limiting environmental conditions. SO2 absorption quantities are then used to predict relative photosynthesis following the fumigation. Thus, relative photosynthesis of plants following fumigation can be predicted on the basis of conductance in SO2-free air. This approach to predicting SO2 resistances of plants includes analysis of their stomatal responses to fumigation, their characteristics of SO2 adsorption and absorption, and their change in photosynthesis resulting from SO2 stress.


Supramolecular isomerism, single-crystal to single-crystal transformation induced by release of in situ generated I2 between two supramolecular frameworks.
Two supramolecular isomers, [H2tpim·H4betc]·I (1) and [H2tpim·H4betc]·I3·2H2O (2), have been synthesized by one-pot reactions. Single-crystal to single-crystal transformations are observed between the two supramolecular isomers which are induced by release of in situ generated I2.


Treatment with A2A receptor antagonist KW6002 and caffeine intake regulate microglia reactivity and protect retina against transient ischemic damage.
Transient retinal ischemia is a major complication of retinal degenerative diseases and contributes to visual impairment and blindness. Evidences indicate that microglia-mediated neuroinflammation has a key role in the neurodegenerative process, prompting the hypothesis that the control of microglia reactivity may afford neuroprotection to the retina against the damage induced by ischemia-reperfusion (I-R). The available therapeutic strategies for retinal degenerative diseases have limited potential, but the blockade of adenosine A2A receptor (A2AR) emerges as candidate strategy. Therefore, we evaluated the therapeutic potential of a selective A2AR antagonist (KW6002) against the damage elicited by I-R. The administration of KW6002 after I-R injury reduced microglia reactivity and inflammatory response and afforded protection to the retina. Moreover, we tested the ability of caffeine, an adenosine receptor antagonist, in mediating protection to the retina in the I-R injury model. We demonstrated that caffeine administration dually regulated microglia reactivity and cell death in the transient retinal ischemic model, depending on the reperfusion time. At 24 h of reperfusion, caffeine increased microglial reactivity, inflammatory response and cell death elicited by I-R. However, at 7 days of reperfusion, caffeine administration decreased microglia reactivity and reduced the levels of proinflammatory cytokines and cell death. Together, these results provide a novel evidence for the use of adenosine A2AR antagonists as potential therapy for retinal ischemic diseases and demonstrate the effect of caffeine on the regulation of microglia-mediated neuroinflammation in the transient ischemic model.


Ictal smile.
Smiling is sometimes manifested during partial seizures. Its value for localizing the epileptogenic focus is not known. We analyzed smiling as an ictal manifestation possibly useful for seizure localization. We reviewed patients referred to the video-EEG monitoring unit who presented a smile as part of their critical symptoms. Ictal smile was defined as an accordant expression accompanied by other characteristic epileptic symptoms and ictal EEG activity. Five of 86 patients experienced partial seizures with an ictal smile. We observed smiling during parietal (two patients) and temporal lobe (three patients) seizures. The right hemisphere appeared to be involved with greater frequency. Ictal smile is an uncommon manifestation of partial seizures involving temporal or parietal lobes, localized mainly on the right hemisphere.


Erratum to Rate of early onset Alzheimer's disease: a systematic review and meta-analysis.
[This corrects the article on p. 38 in vol. 3, PMID: 25815299.].


Estimating species richness using the jackknife procedure.
An exact expression is given for the jackknife estimate of the number of species in a community and for the variance of this number when quadrat sampling procedures are used. The jackknife estimate is a function of the number of species that occur in one and only one quadrat. The variance of the number of species can be constructed, as can approximate two-sided confidence intervals. The behavior of the jackknife estimate, as affected by quadrat size, sample size and sampling area, is investigated by simulation.


Dietary supplementation of microalgae mitigates the negative effects of heat stress in broilers.
Heat stress in poultry is a serious concern, affecting their health and productivity. To effectively address the issue of heat stress, it is essential to include antioxidant-rich compounds in the poultry diet to ensure the proper functioning of the redox system. Microalgae (Spirulina platensis) are rich in antioxidants and have several health benefits in humans and animals. However, its role in health and production and the underlying mechanism in heat-stressed broilers are poorly understood. This study aimed to determine the effect of microalgae supplementation on the health and production of heat-stressed broilers. Cobb500 day-old chicks (N = 144) were raised in litter floor pens (6 pens/treatment and 8 birds/pen). The treatment groups were: 1) no heat stress (NHS), 2) heat stress (HS), and 3) heat stress + 3% microalgae (HS+MAG). The broilers in the HS+MAG group were fed a diet supplemented with 3% microalgae, whereas NHS and HS groups were fed a standard broiler diet. Broilers in the NHS were raised under standard temperature (20°C-24°C), while HS and HS+MAG broilers were subjected to cyclic heat stress from d 22 to 35 (32°C-33°C for 8 h). Heat stress significantly decreased the final body weight, whereas the supplementation of microalgae increased the final body weight of broilers (P < 0.05). The expressions of ileal antioxidant (GPX3), immune-related (IL4), and tight-junction (CLDN2) genes were increased in microalgae-supplemented broilers compared to heat-stressed broilers (P < 0.05). The ileal villus height to crypt depth ratio was improved in microalgae-supplemented broilers (P < 0.05). In addition, microbial alpha, and beta diversities were higher in the HS+MAG group compared to the HS group (P < 0.05). There was an increase in volatile fatty acid-producing bacteria at the genus level, such as Ruminococcus, Ocillospira, Lactobacillus, Oscillobacter, Flavonifractor, and Colidextribacter in the group that received microalgae supplementation. In conclusion, dietary supplementation of microalgae improved the growth performances of heat-stressed broilers by improving their physiogenomics. Thus, the dietary inclusion of microalgae can potentially mitigate heat stress in broilers.


Relationship between inflammatory biomarker galectin-3 and hippocampal volume in a community study.
Galectin-3 (Gal3) is expressed by microglia and performs functions including adhesion; activation of macrophages and fibroblasts, and mediates inflammatory responses in the hippocampus. The present study examined whether serum Gal3 levels predict hippocampal volume in a multi-ethnic, community-based sample. Results of a multiple linear regression (controlling for depression, serum creatinine level, age, BMI, total brain volume, MoCA score, sex, ethnicity, smoking status, history of diabetes) showed that Gal3 levels significantly predicted left (p = .027) but not right hippocampal volume. The relationship was stronger in men than women. Findings suggest this novel inflammatory biomarker is associated with human hippocampal volume.


Does Gender Nonconforming Behavior in Early Childhood Predict Adolescents' Depressive Symptoms?
Gender nonconforming behavior (GNB) is a risk factor for poorer psychological adjustment, but little is known about whether preschool-age children displaying GNB are at risk for depressive symptoms during adolescence. We examined maternal report of GNB at age 4-5 years-old as a predictor of adolescents' depressive symptoms at age 16-17 years-old in a longitudinal study of U.S. children from a predominantly low SES (61% received Aid to Families with Dependent Children) and African American (90%) sample. Youth with GNB in early childhood (n = 10) reported more depressive symptoms during adolescence than did their peers without GNB (n = 115), and this relationship remained after controlling for covariates (environmental risk, prenatal exposure, and neonatal medical problems). Our findings suggest that early GNB may be a risk factor for the development of depressive symptoms in adolescence. Further research is needed to replicate the current findings with a larger sample and to identify the underlying mechanisms by which GNB may increase risk for depressive symptoms. If replicated, the findings further highlight the need for both professionals and parents to become aware of the potential challenges that children with GNB face and to become knowledgeable about ways to facilitate healthy adjustment among gender nonconforming youth.


Crafting a Profession: Moving From Innovative Model Programs to a Successful National Profession.
The authors review the historical underpinnings of the physician assistant (PA) profession and the tumultuous social context at the time the PA profession was born. They explore the creation of "model" PA programs and the subsequent crafting of the PA profession through 2 distinct "quality control" procedures: the credentialing of PA programs (accreditation) and the credentialing of PA graduates (certification). These pillars of PA education and PA practice brought credibility to a fledgling profession in its early years and stand today as examples of the creative thinking and courage of the architects of a profession that, for the first time in American medical history, allowed someone, other than physicians, to practice medicine.


Chlorohydrin formation from unsaturated fatty acids reacted with hypochlorous acid.
Stimulated neutrophils produce hypochlorous acid (HOCl) via the myeloperoxidase-catalyzed reaction of hydrogen peroxide with chloride. The reactions of HOCl with oleic, linoleic, and arachidonic acids both as free fatty acids or bound in phosphatidylcholine have been studied. The products were identified by gas chromatography-mass spectrometry of the methylated and trimethylsilylated derivatives. Oleic acid was converted to the two 9,10-chlorohydrin isomers in near stoichiometric yield. Linoleic acid, at low HOCl:fatty acid ratios, yielded predominantly a mixture of the four possible monochlorohydrin isomers. Bischlorohydrins were also formed, in increasing amounts at higher HOCl concentrations. Arachidonic acid gave a complex mixture of mono- and bischlorohydrins, the relative proportions depending on the amount of HOCl added. Linoleic acid appears to be slightly more reactive than oleic acid with HOCl. Reactions of oleic and linoleic acids with myeloperoxidase, hydrogen peroxide, and chloride gave chlorohydrin products identical to those with HOCl. Lipid chlorohydrins have received little attention as products of reactions of neutrophil oxidants. They are more polar than the parent fatty acids, and if formed in cell membranes could cause disruption to membrane structure. Since cellular targets for HOCl appear to be membrane constituents, chlorohydrin formation from unsaturated lipids could be significant in neutrophil-mediated cytotoxicity.


Heart transplantation in patients bridged with mechanical circulatory support: outcome comparison with matched controls.
Due to the shortage of heart donors, increasing numbers of heart transplantation (HTx) candidates are receiving long-term mechanical circulatory support (MCS) as bridge-to-transplantation. Treatment with MCS is associated with increased formation of anti-human leukocyte antigen antibodies (allosensitization), but whether this affects post-HTx outcomes is unclear. We included all adult patients who received long-term MCS as bridge-to-transplantation and underwent subsequent HTx at our centre between 2008 and 2018. We also enrolled medically treated HTx recipients without prior MCS as controls. These controls were matched by age, sex, diagnosis, and transplantation era. Outcome parameters were compared between the two study groups. A total of 126 patients (48 ± 15 years, 84% male) were included of whom 64 were bridged with MCS and 62 were matched controls. Pre-HTx allosensitization occurred more frequently in the MCS group than in the control group (27% vs. 11%, P = 0.03). At post-HTx year 10, the overall survival probability was 84% among patients treated with MCS and 90% among those medically managed (P = 0.32). At post-HTx year 1, freedom from treated rejections (≥ISHLT 2R) was 69% in the MCS group and 70% in the control group (P = 0.94); and freedom from any rejection was 8% and 5%, respectively (P = 0.98). There were no differences in renal function or cardiac allograft vasculopathy (grade ≥ 1) between groups at 1, 3, and 5 years post-HTx. Although patients treated with MCS had a higher frequency of pre-HTx allosensitization, there were no significant differences in post-HTx graft survival, biopsy-proven rejections, or renal function as compared with patients not bridged with MCS.


Advanced maternal age as a risk factor for stress urinary incontinence: a review of the literature.
The pathophysiology of stress urinary incontinence (SUI) is multifactorial and evidence supports a critical role of pregnancy and vaginal delivery. This review dissects epidemiologic literature to determine the weight of evidence on the role of advanced maternal age (AMA) as a risk factor for the development of subsequent or persistent SUI. We conducted a Medline search using the keywords postpartum, SUI, maternal age, pregnancy, and incontinence. The published literature was critically analyzed. Evidence supports that childbirth trauma contributes to the development and severity of SUI. Yet, there is contradicting evidence as to whether AMA increases the risk. AMA clearly represents an independent risk factor for postpartum SUI. However, long-term studies did not confirm this observation. Whether this finding is suggestive of a true biologic signal that is lost with competing risk factors over time warrants further research.


Prognostic factors in chronic lymphocytic leukaemia: the importance of age, sex and response to treatment in survival. A report from the MRC CLL 1 trial. MRC Working Party on Leukaemia in Adults.
We report the analysis of prognostic factors in a cohort of 660 patients entered in the first Medical Research Council trial in chronic lymphocytic leukaemia (CLL) between 1978 and 1984. The majority (94%) of patients were aged 50 or over and the number of men (M) was almost twice that of women (F) with an M:F ratio of 1.8:1. The M:F ratio was lower, 1.5:1, in patients aged 70 or over. Stage A CLL was the most common, and stage C the least common, among women of all ages, in contrast to men for whom stage A only predominated in the older age group. As the majority of CLL patients are elderly we have examined the causes of death in great detail. 29% of deaths were unrelated to CLL, mainly other cancers (12%) and cardiovascular complications (16%). The majority of deaths in patients presenting with stages B and C were from CLL-related causes, whilst almost half of the deaths in patients presenting with stage A were not obviously related to CLL. Univariate analysis disclosed that the A, B, C staging system was the most important factor considered; stratified and multivariate analysis showed that age and response to treatment were the main prognostic factors after stage. Women always fared better than men and this was independent of stage and age. This and other features documented in the trial suggest a major biological difference between the sexes which has not been widely recognized. The significant influence of treatment response on patients' survival suggests that the search for better treatments in CLL may be rewarding. The improved median survival of stage C patients recorded in this trial, 41 months, compares favourably with previous reports and may have resulted from better treatment.


Inhibition by antiserum to vasoactive intestinal polypeptide (VIP) of prolactin secretion induced by serotonin in the rat.
Intraventricular (i.c.v.) injection of serotonin (5HT) or intravenous (i.v.) injection of 5-hydroxytryptophan (5HTP), a precursor of 5HT, raised plasma prolactin (PRL) levels in urethane-anesthetized rats pretreated with normal rabbit serum. When the animals were injected i.c.v. or i.v. with specific anti-VIP rabbit serum, the plasma PRL responses to 5HT and 5HTP were blunted. These findings suggest that hypothalamic VIP is involved, at least in part, in PRL secretion induced by central serotonergic stimulation in the rat.


Discovery of novel brain-penetrant GluN2B NMDAR antagonists via pharmacophore-merging strategy as anti-stroke therapeutic agents.
In this work, a novel structural series of brain-penetrant GluN2B NMDAR antagonists were designed, synthesized and biologically evaluated as anti-stroke therapeutic agents via merging the structures of NBP and known GluN2B ligands. Approximately half of them exhibited superior neuroprotective activity to NBP against NMDA-induced neurotoxicity in hippocampal neurons at 10 μM, and compound 45e and 45f exerted equipotent activity to ifenprodil, an approved GluN2B- selective NMDAR antagonist. In particular, 45e, with the most potent neuroprotective activity throughout this series, displayed dramatically enhanced activity (Ki = 3.26 nM) compared to ifenprodil (Ki = 14.80 nM) in Radioligand Competitive Binding Assay, and remarkable inhibition (IC50 = 79.32 nM) against GluN1/GluN2B receptor-mediated current in Patch Clamp Assay. Meanwhile, 45e and its enantiomers exhibited low inhibition rate against the current mediated by other investigated receptors at the concentration of 10 μM, indicating their favorable selectivity for GluN1/GluN2B. In the rat model of middle cerebral artery ischemia (MCAO), 45e exerted comparable therapeutic efficacy to ifenprodil at the same dosage. In addition to the attractive in vitro and in vivo potency, 45e displayed a favorable bioavailability (F = 63.37%) and an excellent brain exposure. In further repeated dose toxicity experiments, compound 45e demonstrated an acceptable safety profile. With the above merits, 45e is worthy of further functional investigation as a novel anti-stroke therapeutic agent.


The association between circulating levels of antimüllerian hormone and follicle number, androgens, and menstrual cycle characteristics in young women.
To investigate the association between serum antimüllerian hormone (AMH) and other reproductive parameters in young women. Cross-sectional study. University hospital. Population-based cohort of 256 women: 180 were users and 76 were nonusers of hormonal contraceptives. None. Antral follicles, androgens, age at menarche, and duration and regularity of menstrual cycle. AMH levels were lower among users of hormonal contraceptives compared to nonusers. Among nonusers, women with AMH levels in the upper tertile had 55% (95% confidence interval [CI]: 22%-99%) higher levels of total T and 8% (95% CI = 2%-15%) longer menstrual cycles than women with AMH levels in the lower tertile. An increase of 1 ng/mL in AMH was associated with 45% (95% CI = 6%-97%) higher prevalence of irregular menstrual cycles. These associations were not seen among users of hormonal contraceptives. A strong relationship between AMH and follicle number was found in both users and nonusers. AMH measurements were found to be applicable in evaluation of the reproductive function of young women. However, there may be differences in the way that serum AMH levels can be interpreted depending on whether the woman uses hormonal contraceptives or not.


A rapid staining technique for the detection of the initiation of germination of bacterial spores.
We propose to apply the Wirtz-Conklin staining technique to evaluate spore germination. Spores at different stages of germination were stained with modified spore stain (Wirtz-Conklin) and evaluated for staining properties. Bacillus spores suspended in deionized water, which does not support germination, stained greenish-blue. Spores suspended in germination enhancers that did not form bacilli stained pink, indicating the initiation of germination. Spores suspended in culture media, which promotes bacterial outgrowth, formed bacilli and were also stained pink. Modified spore stain (Wirtz-Conklin) was found to be useful to detect the initiation of spore germination as early as 30 min following incubation in a germination environment. This simple staining procedure is useful in detecting the initiation of germination of bacterial spores.


Clot life span model analysis of clot growth and fibrinolysis in normal subjects: role of thrombin activatable fibrinolysis inhibitor.
Hypofibrinolysis plays a role in thrombophilic states, and a thrombelastography-based method incorporating tissue-type plasminogen activator to determine vulnerability to fibrinolytic stress has recently been developed. This study proposed to kinetically define fibrinolytic vulnerability and the contribution of thrombin activatable fibrinolysis inhibitor to fibrinolytic defenses in normal subjects. Plasma from 30 normal subjects was exposed to tissue factor/kaolin and tissue-type plasminogen activator (100 IU/ml). Prior to activation of coagulation, samples were either not exposed or exposed to potato carboxypeptidase inhibitor (25 microg/ml, a thrombin activatable fibrinolysis inhibitor). Data were collected until clot lysis time was observed. In plasma, time to onset of maximum rate of fibrinolysis was 200-1125 s (95% confidence interval), maximum rate of lysis was -2.0--0.8 dynes/cm2 per s, and clot lysis time was 555-1595 s. Thrombin activatable fibrinolysis inhibitor's inhibition decreased the time to onset of maximum fibrinolysis by 45%, increased the rate of maximum lysis by 50%, and decreased clot lysis time by 45%. The study established a range of fibrinolytic kinetic values and the contribution of thrombin activatable fibrinolysis inhibitor in normal subjects. Study of disease states involving potential hypofibrinolysis (e.g., in-situ ventricular assist device, cancer) could be conducted using this system to link fibrinolytic vulnerability and thrombophilia.


Completely thrombosed giant aneurysm of the pericallosal artery: case report.
Patients with a giant intracranial aneurysm usually present as having an intracranial space-taking lesion, but no subarachnoid hemorrhage. The authors describe a 61-year-old man with a completely thrombosed giant aneurysm at the pericallosal artery. Computed tomography scan showed a large, round, mottled, high-density area in the paramedian frontal region that had ringlike enhancement with contrast media. On cerebral angiography it was demonstrated to be an avascular mass lesion. These findings led to an initial diagnosis of falx meningioma. However, final diagnosis at operation was completely thrombosed giant aneurysm. The literature is reviewed and the pathogenesis of intracranial thrombosed giant aneurysm, in regard to its growth and thrombus formation, is discussed.


A comparison of three balance-assessment scales for patients after stroke with various levels of balance disorder.
It is often difficult for the clinician to choose the most appropriate balance-assessment measure. We wanted to facilitate this decision based on the stroke patient's functional abilities. The aim of our study was to compare three established scales [Berg Balance Scale (BBS), mini-BESTest (MBT) and Functional Gait Assessment (FGA)] in terms of responsiveness, floor and ceiling effects at different levels of ambulation as defined by the Functional Ambulation Classification (FAC). The 18-month prospective study included 88 patients after cerebral stroke, who were able to walk independently or with assistance of one person (FAC 2-6). BBS showed the highest relative gain in the FAC 2-3 group (17% of maximum scale score); in the other two groups (FAC 4-5 and FAC 6), MBT showed the highest relative gain (16 and 13%, respectively), followed by FGA (11 and 10%, respectively). Among the patients with initial FAC 2-3, a floor effect occurred with FGA, while a ceiling effect occurred with BBS among patients with initial FAC 6. Gain in FGA correlated slightly more with improvement detected by MBT (r = 0.60) than with BBS (r = 0.50). We can conclude that BBS seems to be suitable for stroke patients with initial FAC 2-5, whereas MBT and FGA for those with FAC 4-6.


Trapezius upper portion trigger points treatment purpose in positional release therapy with electromyographic analysis.
This quantification process is made through electromyography analysis. This technique of analysis is able to provide a general view of the tension decrease in the superior muscle fibers of the trapezius after therapy. The focus of the present work is to evaluate the treatment of the cervicobrachialgia by Positional Release Therapy (PRT). The present work studies six patients, with ages 44 to 63 (1 male and 5 female) who present tension in the trapezius upper portion fibers. All patients were submitted to 10 session of 30 minutes each. The electromyography was collected on the first and tenth day of treatment. The results demonstrated a progressive decrease of pain in each session. The tension was evaluated by the electromyography analysis, which showed the relations between time of treatment and less pain. With these results, it was possible to verify quantitatively the efficiency of the PRT in the improvement of life quality.


The Rebif new formulation story: it's not trials and error.
The rapid evolution of the biopharmaceutical industry and the development of innovative technologies have provided an opportunity to improve recombinant interferon (IFN)-beta formulations. A number of strategies have been developed to improve the stability, tolerability and immunogenicity of IFNbeta formulations that are used in the long-term treatment of patients with multiple sclerosis (MS). This review focuses on the production of recombinant IFNs and discusses the development of one such biopharmaceutical, Rebif New Formulation (RNF).RNF was developed with the aim of further improving the tolerability and immunogenicity of Rebif, an approved IFNbeta-1a formulation administered subcutaneously three times per week (sc tiw). To this end, numerous candidate drug vehicles and formulations were developed. However, unlike other formulations of IFNbeta, the new candidate formulations in this case were free from all serum-derived components. Specifically, each RNF candidate was free from human serum albumin and produced without fetal bovine serum. The physicochemical stability, injection-site tolerability, pharmacokinetic profile and immunogenic potential of each candidate formulation were systematically tested. This involved initial screening of a large pool of formulations for promising candidates. Two candidate formulations were selected and subjected to further, extensive evaluation.Ex vivo T-cell assays were used to compare the immunogenicity of RNF candidates with that of the current (at the time of writing) approved formulation and an IFNbeta standard. A single RNF candidate induced less T-cell activation, in terms of proliferation and proinflammatory cytokine secretion, than the other two formulations. The results provided ex vivo evidence of the improved immunogenic potential of RNF. A murine model was used to compare the relative immunogenicity of RNF in vivo with two approved formulations of IFNbeta-1a. Mice treated with RNF developed neutralising antibodies more slowly and produced lower titres than mice treated with equivalent doses of the current IFNbeta-1a sc tiw formulation or another approved IFNbeta-1a formulation administered intramuscularly once per week (Avonex). RNF also demonstrated better local tolerability than the current IFNbeta-1a sc tiw formulation after single subcutaneous doses in healthy volunteers. One RNF candidate was superior to the others in all preclinical and phase I studies, and was chosen as the final RNF. This formulation is currently undergoing assessment in a 96-week, phase IIIb clinical trial in patients with MS. This single-arm, open-label, multicentre study will compare the immunogenicity and tolerability of RNF with historical data on the current formulation; results of a 48-week, interim analysis indicate that RNF has improved local tolerability and immunogenicity compared with the current formulation. It is anticipated that the benefits of RNF will translate into an improved long-term benefit-to-risk profile. Further assessment of RNF and other MS drugs is ongoing with the aim of enhancing the therapeutic options available for patients with MS.


Expression of somite segmentation genes in amphioxus: a clock without a wavefront?
In the basal chordate amphioxus (Branchiostoma), somites extend the full length of the body. The anteriormost somites segment during the gastrula and neurula stages from dorsolateral grooves of the archenteron. The remaining ones pinch off, one at a time, from the tail bud. These posterior somites appear to be homologous to those of vertebrates, even though the latter pinch off from the anterior end of bands of presomitic mesoderm rather than directly from the tail bud. To gain insights into the evolution of mesodermal segmentation in chordates, we determined the expression of ten genes in nascent amphioxus somites. Five (Uncx4.1, NeuroD/atonal-related, IrxA, Pcdhdelta2-17/18, and Hey1) are expressed in stripes in the dorsolateral mesoderm at the gastrula stage and in the tail bud while three (Paraxis, Lcx, and Axin) are expressed in the posterior mesendoderm at the gastrula and neurula stages and in the tail bud at later stages. Expression of two genes (Pbx and OligA) suggests roles in the anterior somites that may be unrelated to initial segmentation. Together with previous data, our results indicate that, with the exception that Engrailed is only segmentally expressed in the anterior somites, the genetic mechanisms controlling formation of both the anterior and posterior somites are probably largely identical. Thus, the fundamental pathways for mesodermal segmentation involving Notch-Delta, Wnt/beta-catenin, and Fgf signaling were already in place in the common ancestor of amphioxus and vertebrates although budding of somites from bands of presomitic mesoderm exhibiting waves of expression of Notch, Wnt, and Fgf target genes was likely a vertebrate novelty. Given the conservation of segmentation gene expression between amphioxus and vertebrate somites, we propose that the clock mechanism may have been established in the basal chordate, while the wavefront evolved later in the vertebrate lineage.


Synthesis of dendritic cobalt with flower-like structure by a facile wet chemistry method as an excellent electromagnetic wave absorber.
In this study, a three-dimensional (3D) floral dendritic cobalt (FDC) consisting of layered flakes was effectively synthesized using a facile wet chemistry method. The impact of the molar amount of NaOH on the microscopic morphology, magnetic characteristics, and electromagnetic wave (EMW) absorption properties of the FDC magnetic materials was comprehensively investigated. The results revealed that the prepared FDC features primary, secondary, and multi-level branches, with the majority of secondary branches being parallel to one another. The dendrites grew closely towards the flower's center at one end, while the tips extend in various directions, forming a dendritic flower cluster. The optimal reflection loss (RL) of S3 at 9.3 GHz was -56.34 dB with a thickness of 1.89 mm, and the maximum effective absorption bandwidth (EAB, RL < -10 dB) reached 6.0 GHz (12.0-18.0 GHz) at a thickness of 1.30 mm. Consequently, the FDC magnetic materials produced in this study presented a method for fabricating high-performance electromagnetic wave absorption (EMWA) materials.


Study on the grafting of chitosan-gelatin microcapsules onto cotton fabrics and its antibacterial effect.
In this work, patchouli oil embedded chitosan-gelatin microcapsules were prepared by the complex coacervation method. Their surface and morphology were characterized by scanning electron microscopy (SEM), showing that the microcapsules were with a regular spherical shape in the range of 1 and 20 μm. In the thermal stability analysis, the microcapsules were stable below 190°C, which indicated the fabrics finish can be conducted at 160°C. The encapsulation efficiency (EE) and loading capacity (LC) of the microcapsules were calculated as 50.69% and 30.31%, respectively. Then, the microcapsules were grafted onto cotton fabrics using 2D resin (dimethyloldihydroxyethylene urea, DMDHEU) as the crosslinking reagent. SEM showed that the microcapsules were not only grafted on the surface of the fabrics, but also inserted in the spacing of the fibers. In addition, the formation of ether bonds between 2D resin and hydroxyl groups of cotton and/or hydroxyl groups of the microcapsules was identified by Fourier transform infrared spectroscopy (FTIR). Finally, the antibacterial rate of the fabrics for Staphylococcus aureus and Escherichia coli were about 65% even after being washed 25 times, suggesting its potential application in many fields such as antibacterial mask, bacteriostatic sheet and health-care clothes.


Comparison of the phospholipase activity of bovine milk lipoprotein lipase against rat plasma very low density and high density lipoprotein.
The hydrolytic activity of a lipoprotein lipase from bovine milk against triacylglycerol and phosphatidylcholine of rat plasma very low density lipoprotein was determined and compared to that against phosphatidylcholine of high density lipoprotein. 85--90% of the triacylglycerol in very low density lipoprotein were hydrolyzed to fatty acids and 25--35% of the phosphatidylcholine to lysophosphatidylcholine. High density lipoprotein phosphatidylcholine was only minimally susceptible to the enzyme. Even with high amounts of enzyme and prolonged incubation periods, lysophosphatidylcholine generation did not exceed 2--4% of the original amounts of labeled phosphatidylcholine in the high density lipoprotein. We conclude that phospholipids in high density lipoprotein are not substrates for the phospholipase activity of this lipoprotein lipase. These observations suggest that factors other than the presence of apolipoprotein C-II and of glycerophosphatides are of importance for the activity of lipoprotein lipases.


p53-independent ibrutinib responses in an Eμ-TCL1 mouse model demonstrates efficacy in high-risk CLL.
Deletion of the short-arm of chromosome 17 (17p-) is one of the most critical genetic alterations used in chronic lymphocytic leukemia (CLL) risk stratification. The tumor suppressor TP53 maps to this region, and its loss or mutation accelerates CLL progression, hampers response to chemotherapy and shortens survival. Although florescent in situ hybridization analyses for 17p deletions are routinely performed during clinical diagnoses, p53 mutational status is often unexamined. Given the limited clinical data that exists for frontline treatment of patients with CLL harboring TP53 mutations, there is a need to understand the biology of CLL with TP53 mutations and identify treatment strategies for this subset of patients. Herein, we used a CLL mouse model (Eμ-TCL1) harboring one of the most common TP53 hot-spot mutations observed in CLL (p53(R172H), corresponding to p53(R175H) in humans) to evaluate its impact on disease progression, survival, response to therapy and loss of the remaining wild-type Trp53 allele following ibrutinib treatment. We show that ibrutinib was effective in increasing survival, activating cellular programs outside the p53 pathway and did not place selective pressure on the remaining wild-type Trp53 allele. These data provide evidence that ibrutinib acts as an effective treatment for aggressive forms of CLL with TP53 mutations.


Anchoring genome sequence to chromosomes of the central bearded dragon (Pogona vitticeps) enables reconstruction of ancestral squamate macrochromosomes and identifies sequence content of the Z chromosome.
Squamates (lizards and snakes) are a speciose lineage of reptiles displaying considerable karyotypic diversity, particularly among lizards. Understanding the evolution of this diversity requires comparison of genome organisation between species. Although the genomes of several squamate species have now been sequenced, only the green anole lizard has any sequence anchored to chromosomes. There is only limited gene mapping data available for five other squamates. This makes it difficult to reconstruct the events that have led to extant squamate karyotypic diversity. The purpose of this study was to anchor the recently sequenced central bearded dragon (Pogona vitticeps) genome to chromosomes to trace the evolution of squamate chromosomes. Assigning sequence to sex chromosomes was of particular interest for identifying candidate sex determining genes. By using two different approaches to map conserved blocks of genes, we were able to anchor approximately 42 % of the dragon genome sequence to chromosomes. We constructed detailed comparative maps between dragon, anole and chicken genomes, and where possible, made broader comparisons across Squamata using cytogenetic mapping information for five other species. We show that squamate macrochromosomes are relatively well conserved between species, supporting findings from previous molecular cytogenetic studies. Macrochromosome diversity between members of the Toxicofera clade has been generated by intrachromosomal, and a small number of interchromosomal, rearrangements. We reconstructed the ancestral squamate macrochromosomes by drawing upon comparative cytogenetic mapping data from seven squamate species and propose the events leading to the arrangements observed in representative species. In addition, we assigned over 8 Mbp of sequence containing 219 genes to the Z chromosome, providing a list of genes to begin testing as candidate sex determining genes. Anchoring of the dragon genome has provided substantial insight into the evolution of squamate genomes, enabling us to reconstruct ancestral macrochromosome arrangements at key positions in the squamate phylogeny, demonstrating that fusions between macrochromosomes or fusions of macrochromosomes and microchromosomes, have played an important role during the evolution of squamate genomes. Assigning sequence to the sex chromosomes has identified NR5A1 as a promising candidate sex determining gene in the dragon.


Comparative study of the effects of amphotericin B on the glucose metabolism in Saccharomyces cerevisiae in K(+)- and Na(+)-rich media.
In order to elucidate the effects of amphotericin B (AMB) on the glycolytic pathway, the metabolism of [1-13C]glucose in glucose-grown repressed Saccharomyces cerevisiae was studied. The cells were aerobically suspended in pyrophosphate solutions of high potassium concentration with or without 10(-6) M amphotericin B and measurements were made using 1H-, 13C-NMR spectroscopy and biochemical methods. The results were compared with those obtained under the same experimental conditions but in a medium rich in sodium salts containing the same antibiotic concentration. In general the presence of 10(-6) M AMB reduces the glucose consumption and the ethanol production while favouring the glycerol and trehalose formation. These effects are greatly reduced when a high K+ concentration was used. The AMB effects on the glucose consumption and the production of ethanol, glycerol and trehalose, observed in a suspension rich in Na+, can be fairly well explained by the leakage of K+ through AMB membrane channels. This outflux induces a substantial decrease in the activity of some K(+)-dependent enzymes, such as aldolase, phosphofructokinase and pyruvate kinase. The intensities of the glutamate C2 and C4 signals are higher with a suspension rich in Na+ than with a suspension rich in K+, suggesting that the Krebs cycle operates more effectively in a solution rich in Na+. In the absence of AMB, the passive diffusion of glycerol through the cell membrane is relatively slow and apparently depends on the ionic external medium: it is more efficient in solutions with a high K+ than with a high Na+ concentration. In the presence of 10(-6) M AMB, the glycerol C1,3 resonance drastically decreases at 20 min and then disappears in the noise. This rapid disappearance suggests that glycerol can easily pass through the pores arising from the interaction of AMB with the membrane sterols. However, the rate of pore formation is slow, independent of the external medium (Na+ or K+) and this process is not completed within 20 min.


Daidzein ameliorates diabetic retinopathy in experimental animals.
The present study was designed to check the effect of daidzein in the management of diabetic retinopathy. Streptozotocin at dose 55 mg/kg was used for inducing diabetes in rats. After 28 days of diabetic induction, animals were treated with daidzein at dose 25, 50, and 100 mg/kg for the next 28 days. Electroretinography, estimation of plasma glucose, lactate dehydrogenase, aldose reductase, sorbitol dehydrogenase and oxidative stress parameters were performed at the end of the study. Histopathology of retina was carried out at the end of the study. Diabetic control animals showed a significant increase in levels of plasma glucose and plasma lactate dehydrogenase (p < 0.001). Treatment with daidzein at a dose of 50 and 100 mg/kg significantly reduced the elevated level of blood glucose (p < 0.01 and p < 0.01). Whereas, treatment with daidzein at a dose 100 mg/kg significantly reduced the elevated level of lactate dehydrogenase in plasma after 28 days of treatment (p < 0.01). Treatment with daidzein at a dose of 100 mg/kg significantly reduced the level of aldose reductase and sorbitol dehydrogenase (p < 0.01 and p < 0.001 respectively). Electroretinography revealed that daidzein treatment at a dose of 100 mg/kg significantly prevented the change in 'a' and 'b' wave amplitude and latency. Oxidative stress was also found to be significantly reduced after 28 days of daidzein treatment. Histopathological findings showed a reduction in retinal thickness after daidzein treatment. Daidzein treatment protected retina from damage in hyperglycaemic conditions. Thus, Daidzein can be considered as an effective treatment option for diabetic retinopathy.


Education of military veterans in physician assistant programs.
To assess the admission policies, experiences, and attitudes of physician assistant (PA) program directors regarding recruiting, admitting, and training military veterans after the announcement of the "Helping Veterans Become Physician Assistants" initiative. A descriptive survey of 22 questions was distributed to all 173 PA program directors in the United States in April 2013. The survey covered years 2011 to 2013, although it was completed in June of 2013. The results of the survey were compared with the results of a similar survey that covered years 2008 through 2010. One hundred and five (60.7%) program directors participated in this survey. Veterans were admitted into 88.1% of responding programs and accounted for an average of 4.0% of all students. One-third of programs (33%) accepted transfer credits for veterans' military training, and 20% accepted credits for off-duty education. One-third (33%) of programs participated in the Yellow Ribbon Program. Almost 60% of programs had military veteran faculty members. Active recruitment of military veteran students occurred in 31.2% of programs. Program directors described multiple benefits of, and barriers to, admitting and educating veterans. For the years 2011 through 2013, as compared to 2008 through 2010, there was an increase in the percentage of PA programs that actively recruited veterans, considered veteran status in the admission process, admitted veterans, and contributed to their financial support. There was also an increased percentage of students with military experience matriculating into PA programs. However, barriers still exist for veterans seeking admission into PA programs, the most significant of which is academic preparedness for a graduate-level PA program.


The infra-isthmal fracture of the shaft of the femur.
The common infra-isthmal fracture of the shaft of the femur is defined. The difficulties in its management are discussed and various methods of treatment are critically reviewed. The operative treatment of 21 such fractures with a single plate by the AO method is described. Strict adherence to every detail of the operative and postoperative technique and a cooperative patient are essential for a successful result.


Determination of the handedness of the crossbridge helix of Limulus thick filaments.
Thick filaments, isolated in their long conformation from unstimulated Limulus telson muscles, were shadowed with platinum or platinum-carbon and examined using electron microscopy and optical diffraction techniques. All filaments showed evidence of a right-handed surface helix, which had a major repeat at approx. 43 nm. In fortuitously oriented specimens the subunits, presumably crossbridges, which comprised the helical strands were clearly delineated. Optical transforms obtained from images of shadowed filaments confirmed the helical repeat at approx. 43 nm and could be readily interpreted as patterns expected from a one-surface view of the four-stranded filament structure we have previously reported. The striking resemblance between optically filtered images of shadowed filaments and the computed reconstruction of the one-surface filament further confirm our model for the myosin lattice of the Limulus thick filament.


Lowest excited triplet states of hypericin and isohypericin.
The fluorescence and phosphorescence of hypericin and isohypericin were studied in an ethanol matrix at 1.2 K. The prompt fluorescence spectra are mirror images of the absorption around the 0-0 transition, as expected. The 0-0 vibronic lines of the phosphorescence are found at 13,190 and 13,622 cm-1, and the phosphorescence decay times are 2.79 and 6.6 ms at 1.2 K for hypericin and isohypericin respectively. The fluorescence excitation spectrum of isohypericin reveals a small trace of a different pigment, possibly a tautomeric form of isohypericin. The decay of the phosphorescence of isohypericin is biexponential. The slow component is attributed to an unknown contaminant with a high quantum yield of phosphorescence.


The Response of Acinetobacter baumannii to Zinc Starvation.
Zinc (Zn) is an essential metal that vertebrates sequester from pathogens to protect against infection. Investigating the opportunistic pathogen Acinetobacter baumannii's response to Zn starvation, we identified a putative Zn metallochaperone, ZigA, which binds Zn and is required for bacterial growth under Zn-limiting conditions and for disseminated infection in mice. ZigA is encoded adjacent to the histidine (His) utilization (Hut) system. The His ammonia-lyase HutH binds Zn very tightly only in the presence of high His and makes Zn bioavailable through His catabolism. The released Zn enables A. baumannii to combat host-imposed Zn starvation. These results demonstrate that A. baumannii employs several mechanisms to ensure bioavailability of Zn during infection, with ZigA functioning predominately during Zn starvation, but HutH operating in both Zn-deplete and -replete conditions to mobilize a labile His-Zn pool.


Bisphosphonates for cancer treatment: Mechanisms of action and lessons from clinical trials.
A growing body of evidence points toward an important anti-cancer effect of bisphosphonates, a group of inexpensive, safe, potent, and long-term stable pharmacologicals that are widely used as osteoporosis drugs. To date, they are already used in the prevention of complications of bone metastases. Because the bisphosphonates can also reduce mortality in among other multiple myeloma, breast, and prostate cancer patients, they are now thoroughly studied in oncology. In particular, the more potent nitrogen-containing bisphosphonates have the potential to improve prognosis. The first part of this review will elaborate on the direct and indirect anti-tumoral effects of bisphosphonates, including induction of tumor cell apoptosis, inhibition of tumor cell adhesion and invasion, anti-angiogenesis, synergism with anti-neoplastic drugs, and enhancement of immune surveillance (e.g., through activation of γδ T cells and targeting macrophages). In the second part, we shed light on the current clinical position of bisphosphonates in the treatment of hematological and solid malignancies, as well as on ongoing and completed clinical trials investigating the therapeutic effect of bisphosphonates in cancer. Based on these recent data, the role of bisphosphonates is expected to further expand in the near future outside the field of osteoporosis and to open up new avenues in the treatment of malignancies.


Naturally occurring diacetyl and 2,3-pentanedione concentrations associated with roasting and grinding unflavored coffee beans in a commercial setting.
Over the last decade, concerns have been raised about potential respiratory health effects associated with occupational exposure to the flavoring additives diacetyl and 2,3-pentanedione. Both of these diketones are also natural components of many foods and beverages, including roasted coffee. To date, there are no published studies characterizing workplace exposures to these diketones during commercial roasting and grinding of unflavored coffee beans. In this study, we measured naturally occurring diacetyl, 2,3-pentanedione, and respirable dust at a facility that roasts and grinds coffee beans with no added flavoring agents. Sampling was conducted over the course of three roasting batches and three grinding batches at varying distances from a commercial roaster and grinder. The three batches consisted of lightly roasted soft beans, lightly roasted hard beans, and dark roasted hard beans. Roasting occurred for 37 to 41 min, and the grinding process took between 8 and 11 min. Diacetyl, 2,3-pentanedione, and respirable dust concentrations measured during roasting ranged from less than the limit of detection (<LOD) to 0.0039 ppm, <LOD to 0.018 ppm, and <LOD to 0.31 mg/m3, respectively. During grinding, diacetyl, 2,3-pentanedione, and respirable dust concentrations ranged from 0.018 to 0.39 ppm, 0.0089 to 0.21 ppm, and <LOD to 1.7 mg/m3, respectively. For any given bean/roast combination and sample location, diketone concentrations during grinding were higher than those measured during roasting. During grinding, concentrations decreased with increased distance from the source. Measured concentrations of both diketones were higher during grinding of soft beans than hard beans. The results indicate that airborne concentrations of naturally occurring diacetyl and 2,3-pentanedione associated with unflavored coffee processing: (1) are similar to the concentrations that have been measured in food flavoring facilities; (2) are likely to exceed some recommended short-term occupational exposure limits, but; (3) based on previous analyses of exposure response relationships in animal studies, are far below the concentrations that are expected to cause even minimal responses in the human respiratory tract.


The contribution of laparoscopy to the treatment of gastric cancer.
Laparoscopy plays a role in the preoperative diagnosis of gastric cancer, particularly in determining the location and extent of the neoplasia. In addition to its use in staging, laparoscopy is indicated for the gastric resection of T1-T2, and its middle- and long-term results are comparable to those obtainable with open surgery. Herein we describe our experience with the laparoscopic resection of gastric carcinomas, including the dissection of lymph nodes and the Billroth II reconstruction of digestive continuity with gastrojejunostomia. We carried out laparoscopic gastric resections in 25 patients with adenocarcinomas. Our method involved installing five trocars, tying the left and right gastric vessels and the right gastro-epiploic vessels, sectioning the duodenum 3 cm from the pylorus, sectioning the remaining portion of the stomach obliquely 3 cm from the cardias, and performing Billroth II reconstruction. The average duration of the operation was 4 h 45 min. The average number of removed lymph nodes was 30.5 (range, 22-41). Five patients were converted to laparotomy. Significant complications were observed in four cases (16%). Hospitalization ranged from 5 to 16 days. The average follow-up was 38 months (range, 7-63), without evidence of relapse. In terms of morbidity, our results were similar to those obtained with open surgery. Lymphectomy according to the extent and number of lymph nodes is acceptable in the treatment of tumors of the lower third of the stomach. More case studies are needed to provide further indications of the applicability of the technique (which is currently used only in a few centers) and long-term results.


Subclass characteristics of IgG autoantibodies in bullous pemphigoid and pemphigus.
Immunoglobulin (Ig) G subclasses in anti-basement membrane zone (BMZ) autoantibodies found in the sera of bullous pemphigoid (BP) and in anti-intercellular substance (ICS) autoantibodies of pemphigus were investigated using immunofluorescent (IF) staining. In BP, IgG4, IgG1, and IgG2 were detected in 13, 5 and 6 of 15 patients, respectively; IgG3 was not detected. In pemphigus, IgG4 was detected in all of 10 patients, IgG1 in 7, IgG2 in one, and IgG3 in one patient, respectively. In both BP and pemphigus, the most prominent subclass in intensity of IF staining was IgG4. Although one BP and one PV patient had only IgG4 autoantibodies, C3 deposition was detected. The quantification of IgG subclasses in the sera of the patients was performed by enzyme-linked immunosorbent assays (ELISA). Serum levels of IgG4 in both BP and pemphigus were elevated approximately 3-fold over those in normal controls; those of whole IgG and IgG1-3 were not significantly elevated. Using direct IF staining, the deposition of C3 at the BMZ and at the ICS was demonstrated in 9 of 10 BP and in 3 of 8 pemphigus patients, respectively. The prominent IgG subclasses of anti-BMZ and anti-ICS antibody were IgG4, a noncomplement-fixing antibody, suggesting that the deposition of C3 in the lesional skin occurred via the alternative pathway, or that small amounts of IgG1-3 subclass autoantibodies activated the classical pathway.