Clinical Variants of the Native Class D β-Lactamase of Acinetobacter baumannii Pose an Emerging Threat through Increased Hydrolytic Activity against Carbapenems.
The threat posed by the chromosomally encoded class D β-lactamase of Acinetobacter baumannii (OXA-51/66) has been unclear, in part because of its relatively low affinity and turnover rate for carbapenems. Several hundred clinical variants of OXA-51/66 have been reported, many with substitutions of active-site residues. We determined the kinetic properties of OXA-66 and five clinical variants with respect to a wide variety of β-lactam substrates. The five variants displayed enhanced activity against carbapenems and in some cases against penicillins, late-generation cephalosporins, and the monobactam aztreonam. Molecular dynamics simulations show that in OXA-66, P130 inhibits the side-chain rotation of I129 and thereby prevents doripenem binding because of steric clash. A single amino acid substitution at this position (P130Q) in the variant OXA-109 greatly enhances the mobility of both I129 and a key active-site tryptophan (W222), thereby facilitating carbapenem binding. This expansion of substrate specificity represents a very worrisome development for the efficacy of β-lactams against this troublesome pathogen.


The primary B cell response to the O/core region of bacterial lipopolysaccharide is restricted to the Ly-1 lineage.
Experiments were performed to test the hypothesis that Ly-1 B cells respond to antigenic challenge with LPS from gram-negative bacteria. To perform these experiments, the splenic fragment culture system for the study of B cell precursors was used. We found that a significant number of anti-O/core, but not anti-lipid A, precursors expressed the lambda L chain. A restriction of the anti-LPS response to Ly-1 B cells was tested using a Ly-1 depletion protocol. We found that the anti-O/core antibody response was restricted to the Ly-1 B cell lineage. In contrast, conventional B cells, not Ly-1 B cells, respond to an antigenic challenge with lipid A. Our results further support the idea that the Ly-1 B cell lineage serves a direct role in protecting against certain bacterial infections.


Invasion biology of Aedes japonicus japonicus (Diptera: Culicidae).
Aedes japonicus japonicus (Theobald) (Diptera: Culicidae) has recently expanded beyond its native range of Japan and Korea into large parts of North America and Central Europe. Population genetic studies begun immediately after the species was detected in North America revealed genetically distinct introductions that subsequently merged, likely contributing to the successful expansion. Interactions, particularly in the larval stage, with other known disease vectors give this invasive subspecies the potential to influence local disease dynamics. Its successful invasion likely does not involve superior direct competitive abilities, but it is associated with the use of diverse larval habitats and a cold tolerance that allows an expanded seasonal activity range in temperate climates. We predict a continued but slower expansion of Ae. j. japonicus in North America and a continued rapid expansion into other areas as this mosquito will eventually be considered a permanent resident of much of North America, Europe, Asia, and parts of Hawaii.


A PPAR-gamma agonist protects from radiation-induced intestinal toxicity.
Because of its anti-inflammatory, anti-fibrotic, anti-apoptotic and anti-neoplastic properties, the PPAR-γ agonist rosiglitazone is an interesting drug for investigating for use in the prevention and treatment of radiation-induced intestinal damage. We aimed to evaluate the radioprotective effect of rosiglitazone in a murine model of acute intestinal damage, assessing whether radioprotection is selective for normal tissues or also occurs in tumour cells. Mice were total-body irradiated (12 Gy), with or without rosiglitazone (5 mg/kg/day). After 24 and 72 hours, mice were sacrificed and the jejunum was collected. HT-29 human colon cancer cells were irradiated with a single dose of 2 (1000 cells), 4 (1500 cells) or 6 (2000 cells) Gy, with or without adding rosiglitazone (20 µM) 1 hour before irradiation. HT-29-xenografted CD1 mice were irradiated (16 Gy) with or without rosiglitazone; tumour volumes were measured for 33 days. Rosiglitazone markedly reduced histological signs of altered bowel structures, that is, villi shortening, submucosal thickening, necrotic changes in crypts, oedema, apoptosis, and inflammatory infiltrate induced by irradiation. Rosiglitazone significantly decreased p-NF-kB p65 phosphorylation and TGFβ protein expression at 24 and 72 hours post-irradiation and significantly decreased gene expression of Collagen1, Mmp13, Tnfα and Bax at 24 hours and p53 at 72 hours post-irradiation. Rosiglitazone reduced HT-29 clonogenic survival, but only produced a slight reduction of xenograft tumour growth. Rosiglitazone exerts a protective effect on normal tissues and reduces alterations in bowel structures and inflammation in a radiation-induced bowel toxicity model, without interfering with the radiation effect on HT-29 cancer cells. PPAR-γ agonists should be further investigated for their application in abdominal and pelvic irradiation.


Domestic violence.
Domestic violence is an enormous health care issue that primary care physicians are powerfully suited to address. Rarely, however, do primary care physicians recognize that many of the patients who regularly and routinely present to them are victims of abuse. This article is designed to empower America's front-line doctors to recognize and appropriately treat the many victims of domestic violence who currently suffer from this silent epidemic. The reader will come to understand that it is via relatively simple-to-learn techniques that physicians can significantly impact the health and well-being of their communities.


A Case of Recurrent Pasteurella Bacteremia in an Immunocompetent Patient with No Animal Bite.
BACKGROUND Pasteurella multocida is a gram negative-penicillin sensitive bacterium and is part of normal respiratory microbiota of animals (e.g., cats and dogs) and some birds. Various infections in humans, such as cellulitis, rarely bacteremia, endocarditis, meningitis, and septic arthritis, are a result of domestic cat or dog bites. These infections are rarely seen in an immunocompetent person, without an associated animal bite. CASE REPORT We present a case of refractory Pasteurella multocida bacteremia without any animal bite in an immunocompetent person. CONCLUSIONS Pasteurella multocida bacteremia has been seen in immunocompromised patients and mostly after a cat or dog bite or scratch but might also happen in immunocompetent humans with only pet licking rather than biting, which might increase hospital and emergency department visits or admissions in the future.


Electron microscopical and biochemical characterization of infectious pancreatic necrosis virus.
An electron microscopical and biochemical examination of the properties of infectious pancreatic necrosis virus (IPN) and of its ribonucleic acid (RNA) was made. The buoyant density of IPN in CsCl was found to be 1.33 g/cm(3). Electron microscopical examination of the banded virus revealed structures similar in size (74 nm) and shape to reoviruses but lacking a characteristic inner capsid structure. Polyacrylamide gel electrophoretic analysis of IPN-RNA revealed a single non-segmented component of molecular weight 3.2 x 10(6). Its susceptibility to ribonuclease, base composition, and resistance to thermal denaturation indicated a single-stranded RNA structure. However, its sedimentation behavior (16S) independent of ionic strength in sucrose gradients, partial solubility in 2 m LiCl, and ribonuclease resistance in the presence of Mg(2+) suggest an unusual secondary structure of unknown nature. The accumulated data indicate that IPN virus does not belong to either the picornavirus or reovirus groups and may represent a new group of viruses.


Nitric oxide production in the ventilatory muscles in response to acute resistive loading.
The effect of muscle activation on muscle nitric oxide (NO) production remains controversial. Whereas NO release increases in in vitro activated muscles and in vivo limb muscles, diaphragmatic NO synthase (NOS) activity declines after 3 h of inspiratory resistive loading (IRL). We tested in this study the hypotheses that acute IRL decreases diaphragmatic NO derivatives levels and reduces protein expression of neuronal (nNOS), endothelial (eNOS), and inducible (iNOS) NO synthases, as well as 3-nitrotyrosine formation. Anesthetized, tracheostomized, spontaneously breathing adult rats were subjected to IRL (50% of the maximum inspiratory pressure) for 1, 3, or 6 h. Quietly breathing rats served as controls. After 3 h of IRL, muscle eNOS and nNOS protein levels rose by 80 and 60% of control values, respectively. Whereas eNOS expression did not change any further, nNOS expression reached 550% of control values after 6 h of IRL. Strong iNOS protein expression was detected in the diaphragms after 6 h of IRL. Total NO derivatives levels in the diaphragm declined during IRL as a result of reduction in nitrate, nitrite, and nitrosothiols. Diaphragmatic protein tyrosine nitration decreased in response to IRL, and this reduction was mainly due to reduced tyrosine nitration of enolase and aldolase. We conclude that diaphragmatic NO derivatives levels decline in response to IRL and that the rise in diaphragmatic NOS protein expression may be a compensatory response designed to counterbalance the decline in NOS activity.


Extracorporeal Cardiopulmonary Resuscitation.
Around the world, survival rates after cardiac arrest range between <14% for in-hospital (IHCA) and <10% for outof- hospital cardiac arrest (OHCA). This situation could potentially be improved by using extracorporeal membrane oxygenation (ECMO) during cardiopulmonary resuscitation (CPR), i.e. by extracorporeal cardiopulmonary resuscitation (ECPR). A selective literature search of Pubmed and Embase using the searching string ((ECMO) OR (ECLS)) AND (ECPR)) was carried out in February 2023 to prepare an up-to-date review of published trials comparing the outcomes of ECPR with those of conventional CPR. Out of 573 initial results, 12 studies were included in this review, among them three randomized controlled trials comparing ECPR with CPR, involving a total of 420 patients. The survival rates for ECPR ranged from 20% to 43% for OHCA and 20% to 30.4% for IHCA. Most of the publications were associated with a high degree of bias and a low level of evidence. ECPR can potentially improve survival rates after cardiac arrest compared to conventional CPR when used in experienced, high-volume centers in highly selected patients (young age, initial shockable rhythm, witnessed cardiac arrest, therapy-refractory high-quality CPR). No general recommendation for the use of ECPR can be issued at present.


Whole-Body Metabolism and the Musculoskeletal Impacts of Targeting Activin A and Myostatin in Severe Osteogenesis Imperfecta.
Mutations in the COL1A1 and COL1A2 genes, which encode type I collagen, are present in around 85%-90% of osteogenesis imperfecta (OI) patients. Because type I collagen is the principal protein composition of bones, any changes in its gene sequences or synthesis can severely affect bone structure. As a result, skeletal deformity and bone frailty are defining characteristics of OI. Homozygous oim/oim mice are utilized as models of severe progressive type III OI. Bone adapts to external forces by altering its mass and architecture. Previous attempts to leverage the relationship between muscle and bone involved using a soluble activin receptor type IIB-mFc (sActRIIB-mFc) fusion protein to lower circulating concentrations of activin A and myostatin. These two proteins are part of the TGF-β superfamily that regulate muscle and bone function. While this approach resulted in increased muscle masses and enhanced bone properties, adverse effects emerged due to ligand promiscuity, limiting clinical efficacy and obscuring the precise contributions of myostatin and activin A. In this study, we investigated the musculoskeletal and whole-body metabolism effect of treating 5-week-old wildtype (Wt) and oim/oim mice for 11 weeks with either control antibody (Ctrl-Ab) or monoclonal anti-activin A antibody (ActA-Ab), anti-myostatin antibody (Mstn-Ab), or a combination of ActA-Ab and Mstn-Ab (Combo). We demonstrated that ActA-Ab treatment minimally impacts muscle mass in oim/oim mice, whereas Mstn-Ab and Combo treatments substantially increased muscle mass and overall lean mass regardless of genotype and sex. Further, while no improvements in cortical bone microarchitecture were observed with all treatments, minimal improvements in trabecular bone microarchitecture were observed with the Combo treatment in oim/oim mice. Our findings suggest that individual or combinatorial inhibition of myostatin and activin A alone is insufficient to robustly improve femoral biomechanical and microarchitectural properties in severely affected OI mice. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.


[Surgery for Flail Chest with Titanium Plates after Cardiopulmonary Resuscitation;Report of a Case].
A 74-year-old woman was admitted for the management of nausea with hyponatremia. On the 19th day of hospitalization, the patient developed cardiac arrest and needed cardiopulmonary resuscitation (CPR). CPR was successful but the patient developed multiple rib fractures with flail chest by CPR. Because of persistent dyspnea, surgery for fixing of the fractured ribs was performed using 4 titanium plates, with prompt improvement of the flail chest after the operation.


Governmental policy and personal experiences. The development towards integration of mobility disabled students into regular schools in Scandinavia.
The Scandinavian countries have been among the leading countries with regard to integration into society for persons with disabilities. This article focuses on two intimately related questions concerning the reform: 1) How did the normalization principle, as governmental policy, intend to improve the living-situation for people with disabilities? 2) How was the reform experienced by some disabled persons who attended compulsory school during its introduction? Our sources are: a) central government documents and reports concerning integration of mobility disabled persons in Norway and Sweden; b) 40 life history interviews with mobility disabled persons in Scandinavia born between 1955 and 1965. The interviews are theme centered upon experiences in education and work.


Histopathologic Characterization of Mifepristone-induced Ovarian Toxicity in Cynomolgus Monkeys.
Mifepristone, which is an orally active synthetic steroid with antiprogesterone activity, is known as an ovarian toxicant. Because the available data regarding the histopathologic characteristics of ovarian toxicity in nonhuman primates are limited, the present study was undertaken in order to investigate detailed histopathologic changes accompanying mifepristone-induced ovarian toxicity and its relationship to changes in menstrual cycle and circulating sex steroid hormone. Twenty mg/kg of mifepristone was orally administered daily to 4 cynomolgus monkeys for 2 months. Mifepristone inhibited the cyclic increases in circulating estradiol-17β and progesterone levels with associated absence of menstruation. Histopathologically, the ovary in the treated animals showed follicular phase without changes in the percentage of atretic antral follicles, and reduced endometrial thickness was noted in the uterus. These changes indicated that a certain degree of antral follicle development had been retained in spite of the menstrual cycle having been arrested in mifepristone-treated animals. Our investigation suggested that it is important to perform detailed histopathologic examination of reproductive organs with precise knowledge of the characteristics of each menstrual stage to detect ovarian toxicity in nonhuman primates. Monitoring menstrual signs and circulating sex steroid hormone levels provides additional evidence for the investigation of the mechanism of ovarian toxicity.


Reproductive status affects the feeding ecology and social association patterns of female squirrel monkeys (Saimiri collinsi) in an Amazonian rainforest.
When making foraging decisions, female primates may follow specific behavioral strategies that reflect their reproductive state. Lactation is considered the most energetically costly phase for females, but we argue that gestation is also energetically expensive for squirrel monkeys. In this study, we examined whether female squirrel monkeys (a seasonally breeding primate) in different reproductive phases showed significant differences in their foraging ecology. We sampled two wild groups of squirrel monkeys (Saimiri collinsi) using the focal animal method, during 12 months (June 2014 to May 2015). During this time, we quantified the effect of reproductive state (mating, gestation, and lactation) and sex (females vs. males) on activity budgets, foraging efficiency, dietary composition, and nearest neighbors. We found significant effects of both sex and reproductive phase on the mean proportion of time spent foraging, resting, traveling, and being social. Females consumed more insects than did males at all times; among females, time spent eating prey and fruit varied according to reproductive state. These data suggest that, due to their life history and seasonal breeding, reproduction is a costly activity for female Saimiri, and not only during lactation. Therefore, adopting the appropriate behavioral strategies is essential to reduce energetic deficits in females.


Mychildren's: integration of a personally controlled health record with a tethered patient portal for a pediatric and adolescent population.
Personally controlled health records (PCHRs) and patient portals are increasingly being offered by healthcare institutions, employers, insurance companies and commercial entities to allow patients access to their health information. Both applications offer unique services to provide patients with tools to manage their health. While PCHRs allow users ubiquitous, portable, patient controlled access to their health information, traditional patient portals provide provider-tethered applications allowing patients access, but not control of, certain healthcare information, as well as communication and administrative functions, such as secure messaging, appointment management and prescription refill requests, facilitating care at a specific healthcare facility.We describe our approach for the design, content creation, policy development, and implementation of MyChildren's, a unique web-based application leveraging the advantages of both a provider-tethered patient portal and a PCHR to allow patients and their guardians access to the functionality and convenience of a traditional patient portal, as well as the portability and flexibility of a PCHR.


Expansion of the deletion 13q syndrome phenotype: a case report.
The phenotypic description of deletion 13q syndrome is dependent on the location and size of the deleted segment. At present, the syndrome is divided into 3 groups based on the deletion's location relative to chromosomal band 13q32. Groups 1 (proximal to q32) and 2 (including q32) have shown distinctive phenotypes including mental retardation and growth deficiency, whereas group 3 (q33-34 deletion) is defined by the presence of mental retardation but usually the absence of major malformations. The authors report an 8-year-old Hispanic female with dysmorphic facial features, microcephaly, moderate to severe mental retardation, and uncontrolled epilepsy associated with a terminal 13q33.3 deletion. These features expand the characterization of the phenotype associated with group 3 of the 13q deletion syndrome to include major clinical manifestations. This case report will contribute to more accurate genetic counseling as well as may help identify more individuals with this syndrome.


Immunomodulatory action of eicosanoids and other small molecular weight products of macrophages.
There is an increasing body of evidence that T cell-mediated immune response is regulated by a variety of small molecular weight products of macrophages. One of the best known immunoregulatory products in this context is prostaglandin E2 (PGE2) which is known to regulate both T cell and macrophage functions. Recently, ornithine, cysteine, and lactate have also been recognized as immunoregulatory mediators. In analogy to the hormone-like cytokines and lymphokines, all these substances are produced by immunologically relevant cells (macrophages) at a variable and regulated rate; and they have been shown to regulate the functional activities of other cells (T cells and macrophages). This brief review describes the key observations that underscore the important regulatory role of these metabolites in physiological and pathological conditions.


OPTICAL COHERENCE TOMOGRAPHY AND OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY PARAMETERS IN PATIENTS WITH PHACOMATOSIS.
To compare the retinal vasculature characteristics between eyes of patients with and without phacomatosis. Case-control observational study with retinal vasculature evaluation by optical coherence tomography and optical coherence tomography angiography of the macula and disk. The study included 80 eyes. Neurofibromatosis Type 1 patients presented with a higher central macular thickness (P = 0.007), a lower optical disk nervous fiber layer (P = 0.006), a lower perimeter, area, and circularity of the foveal avascular zone (P < 0.05), a higher vascular density of macular avascular layer (AMVD) (P = 0.004), and a lower papillary vascular density of superficial capillary plexus (SPVD) (P = 0.048). Patients with tuberous sclerosis presented with an increase in central macular thickness (P = 0.024) and in vascular densities (P < 0.05) [except for macular vascular density of deep capillary plexus (PMVD), AMVD, and SPVD]. Patients with Sturge-Weber syndrome showed a decrease in optical disk nervous fiber layer (P < 0.001), subfoveal choroid thickness (P = 0.011), macular vascular density of superficial capillary plexus (SMVD) (P = 0.036), and SPVD (P < 0.001). Phacomatosis patients showed statistically significant differences of retinal vasculature characteristics, compared to eyes without pathology. Further studies are needed to determine when and if these parameters change with the course of the disease and if they can be used as biomarkers for disease severity or progression.


Horizontal and vertical movements of host-seeking Ixodes pacificus (Acari: Ixodidae) nymphs in a hardwood forest.
The nymph of the western black-legged tick (Ixodes pacificus) is an important bridging vector of the Lyme disease spirochete (Borrelia burgdorferi) to humans in the far-western United States. The previously unknown dispersal capabilities of this life stage were studied in relation to logs, tree trunks, and adjacent leaf-litter areas in a mixed hardwood forest using mark-release-recapture methods. In two spatially and temporally well-spaced trials involving logs, the estimated mean distances that nymphs dispersed ranged from approximately 0.04 to 0.20 m/day on logs vs 0.11 to 0.72 m/day in litter. Prior to recapture in either trial and within the confines of the sampling grids, the greatest estimated dispersal distances by individual nymphs released on logs, and in litter 0.5 m or 1.5 m from logs, were 2.4, 3.0, and 3.0 m, respectively. Nymphs released on logs or litter tended to remain within the same biotopes in which they were freed while host-seeking. In two simultaneous trials involving trunks spaced close-at-hand, nymphs released at the trunk/litter interface on all four aspects collectively dispersed a mean of 0.353 m/day on trunks vs 0.175 m/day in litter. In either trial, the greatest distances that recaptured nymphs climbed trunks, or dispersed in litter in an encircling 3-m grid, were 1.55 m and 2.97 m, respectively. Nymphs ascending trunks did not exhibit a preference for any one aspect, and the B. burgdorferi-infection prevalences in nymphs that climbed trunks (3.2-4.0%) did not differ significantly from those that moved horizontally into litter (10.5-17.6%). We conclude that I. pacificus nymphs use an ambush host-seeking strategy; that they disperse slowly in all biotopes studied; that they usually continue to host-seek in or on whatever substratum they access initially; and that B. burgdorferi-infected nymphs are as likely to move horizontally as vertically when offered a choice.


Fulminant Septic Shock with Pasteurella multocida in a Young Infant: No Bite, Scratch, or Lick!
This is a case of Pasteurella multocida septic shock encountered in a 7-week-old infant without any bites, scratch marks, or history of licks by pet animals (dog and cats in household). The infant required 3 days of vasopressor support and 4 days of mechanical ventilation to achieve normal hemodynamics. This is an unidentified route of transmission and our literature search for this topic discovered reported cases of life-threatening presentation with Pasteurella infections in the absence of a bite or any form of invasive contact with animals. We believe that this is an important public safety message to restrict animal contact of young infants to prevent severe infection.


Mutagenic spectrum resulting from DNA damage by oxygen radicals.
Oxygen free radicals are highly reactive species that damage DNA and cause mutations. We determined the mutagenic spectrum of oxygen free radicals produced by the aerobic incubation of single-stranded M13mp2 DNA with Fe2+. The Fe2(+)-treated DNA was transfected into component Escherichia coli, and mutants within the nonessential lac Z alpha gene for beta-galactosidase were identified by decreased alpha-complementation. The frequency of mutants obtained with 10 microM Fe2+ was 20- to 80-fold greater than that obtained with untreated DNA. Mutagenesis was greater after the host cells were exposed to UV irradiation to induce the SOS "error-prone" response. The ability of catalase, mannitol, and superoxide dismutase to diminish mutagenesis indicates the involvement of oxygen free radicals. The sequence data on 94 of the mutants establish that mutagenesis results primarily from an increase in single-base substitutions. Ninety-four percent of the mutants with detectable changes in nucleotide sequence were single-base substitutions, the most frequent being G----C transversions, followed by C----T transitions and G----T transversions. The clustering of mutations at distinct gene positions suggests that Fe2+/oxygen damage to DNA is nonrandom. This mutational spectrum provides evidence that a multiplicity of DNA lesions produced by oxygen free radicals in vitro are promutagenic and could be a source of spontaneous mutations.


Ruthenium-Catalyzed ortho C-H Borylation of Arylphosphines.
Efficient, phosphine-directed ortho C-H borylation of arylphosphine derivatives was achieved using Ru catalysts for the first time. The reaction is applicable to various tertiary arylphosphine and arylphosphinite derivatives to give (o-borylaryl)phosphorus compounds in high yields. This reaction enables easy access to a variety of functionalized phosphine ligands and ambiphilic phosphine boronate compounds, thus realizing a new late-stage modification of phosphorus compounds.


Acetone-butanol-ethanol (ABE) fermentation in an immobilized cell trickle bed reactor.
Acetone-butanol-ethanol (ABE) fermentation was successfully carried out in an immobilized cell trickle bed reactor. The reactor was composed of two serial columns packed with Clostridium acetobutylicum ATCC 824 entrapped on the surface of natural sponge segments at a cell loading in the range of 2.03-5.56 g dry cells/g sponge. The average cell loading was 3.58 g dry cells/g sponge. Batch experiments indicated that a critical pH above 4.2 is necessary for the initiation of cell growth. One of the media used during continuous experiments consisted of a salt mixture alone and the other a nutrient medium containing a salt mixture with yeast extract and peptone. Effluent pH was controlled by supplying various fractions of the two different types of media. A nutrient medium fraction above 0.6 was crucial for successful fermentation in a trickle bed reactor. The nutrient medium fraction is the ratio of the volume of the nutrient medium to the total volume of nutrient plus salt medium. Supplying nutrient medium to both columns continuously was an effective way to meet both pH and nutrient requirement. A 257-mL reactor could ferment 45 g/L glucose from an initial concentration of 60 g/L glucose at a rate of 70 mL/h. Butanol, acetone, and ethanol concentrations were 8.82, 5.22, and 1.45 g/L, respectively, with a butanol and total solvent yield of 19.4 and 34.1 wt %. Solvent productivity in an immobilized cell trickle bed reactor was 4.2 g/L h, which was 10 times higher than that obtained in a batch fermentation using free cells and 2.76 times higher than that of an immobilized CSTR. If the nutrient medium fraction was below 0.6 and the pH was below 4.2, the system degenerated. Oxygen also contributed to the system degeneration. Upon degeneration, glucose consumption and solvent yield decreased to 30.9 g/L and 23.0 wt %, respectively. The yield of total liquid product (40.0 wt %) and butanol selectivity (60.0 wt %) remained almost constant. Once the cells were degenerated, they could not be recovered.


Unplanned Readmission within 28 Days of Hospital Discharge in a Longitudinal Population-Based Cohort of Older Australian Women.
This study aimed to estimate the incidence of 28-day unplanned readmission among older women, and associated factors. Data were used from the 1921-1926 birth cohort of the Australian Longitudinal Study on Women's Health. Linkage of self-reported survey data with the Admitted Patient Data Collection allowed the identification of hospital admissions for each woman and the corresponding baseline characteristics. The Cox proportional-hazards model was used to identify factors associated with time to unplanned readmission, using SAS software V 9.4. (SAS Institute, Cary, NC, USA). Of 2056 women with index unplanned admission, 363 (17.5%) were readmitted within 28 days of discharge, and of these 229 (11.14%) had unplanned readmission. Among women with unplanned readmission, 24% were for the same condition as for the index hospitalisation. Cardiovascular diseases were the main diagnoses for the index admission and readmission. Unplanned readmission risk was higher if not partnered (hazard ratio (HR) = 1.43, 95% confidence interval (CI): 1.05-1.95), of non-English speaking background (HR = 1.62%, 95% CI: 1.07-2.47), more than three days length of stay on index admission (HR = 1.41%, 95% CI: 1.04-1.90) and one or two of the assessed chronic diseases (HR = 1.68, 95% CI: 1.19-2.36). At least one in ten women had unplanned readmission at some time between ages 75-95 years. Women who are not partnered, not of English-speaking background, with longer hospital stay and those with multi-morbidity, may need further efforts during their stay and on discharge to mitigate unplanned readmission.


Influence of surface finishing on fracture load and failure mode of glass ceramic crowns.
Ceramic restorations often require adjustments using diamond rotary instruments, which damage the glazed surface. The effect of these adjustments on the fracture behavior of these restorations is unclear. The purpose of this in vitro study was to evaluate the influence of induced surface defects on the fracture load and mode of failure of lithium disilicate-based (LDS) glass ceramic restorations. Premolar crowns were obtained from LDS computer-aided design and computer-aided manufacturing blocks (n=60) and glazed. The crowns were bonded to dentin analog dies and divided into 5 groups (n=12), as follows: glaze; abrasion (diamond rotary instrument 2135); abrasion and reglaze; abrasion and polishing (diamond rotary instrument 2135F, 2135 FF, and polishing devices); and polishing. The topography of the crowns was examined by scanning electron microscopy, and roughness was measured. A compressive load (0.5 mm/min) was applied by a piston to the center of the lingual cusp until fracture. The fracture load was recorded and data were statistically analyzed by ANOVA and the Tukey HSD test (α=.05). Fractured crowns were examined to determine the fracture origin. Polishing and/or reglazing resulted in lower roughness than for the abraded group (P<.05), which did not affect the fracture loads (P=.696). Catastrophic fracture with origin at the intaglio surface was the mode of failure for all the crowns. The experiment design successfully submitted the crowns to a clinical stress state, resulting in a clinically relevant failure. Reglazing or polishing were effective in reducing surface defects. Surface treatments had no effect on the immediate catastrophic failure of LDS crowns.


Synthesis, Structure, and Photochemical Behavior of [5]Heli-viologen Isomers.
The syntheses of isomeric helical viologens that have potential applications in supramolecular chemistry and catalysis have been developed. The structures of the molecules and their solid-state packing motifs have been determined by X-ray crystallography. Computational studies demonstrate that the magnitude of their racemization barriers is primarily determined by the identity of the helical scaffold and is insensitive to the placement of the viologen functional group. The isomers are similar in their photophysical behavior but very different in their photochemical behavior.


Interactions between spherical colloids mediated by a liquid crystal: a molecular simulation and mesoscale study.
Monte Carlo simulations and dynamic field theory (DyFT) are used to study the interactions between dilute spherical particles, dispersed in nematic and isotropic phases of a liquid crystal. A recently developed simulation method (expanded ensemble density of states) was used to determine the potential of mean force (PMF) between the two spheres as a function of their separation and size. The PMF was also calculated by a dynamic field theory that describes the evolution of the local tensor order parameter. Both methods reveal an overall attraction between the colloids in the nematic phase; in the isotropic phase, the overall attraction between the colloids is much weaker, whereas the repulsion at short range is stronger. In addition, both methods predict a new topology of the disclination lines, which arises when the particles approach each other. The theory is found to describe the results of simulations remarkably well, down to length scales comparable to the size of the molecules. At separations corresponding to the width of individual molecular layers on the particles' surface, the two methods yield different defect structures. We attribute this difference to the neglect of density inhomogeneities in the DyFT. We also investigate the effects of the size of spherical colloids on their interactions.


Penicilloyl-dextran conjugates as reagents for skin testing in penicillin allergy.
A procedure for implanting amino groups in the dextran molecule is described. The aminated polymer prepared from a dextran sample with a molecular weight of 20,000 was found to contain approximately 16 amino groups per molecule. Similarly to polylysine, the aminated dextran may be penicilloylated by incubation with penicillin at high pH. The penicilloyl-dextran (P-Dex) conjugate prepared in this way was found to contain approximately 8 penicilloyl (PO) groups per molecule. P-Dex was found practically nonimmunogenic in rabbits and showed itself antigenically active in precipitin, passive hemagglutination and passive cutaneous anaphylaxis (PCA) tests performed with a PO specific antiserum. A good correlation was found between the skin tests performed parallelly with P-Dex and penicilloyl-polylysine (PPL) in four subjects with penicillin allergy.


Association between social adaptability index and survival of patients with chronic kidney disease.
While individual socioeconomic factors have been associated with clinical outcome, a composite index has not been developed. In this project, we tested the hypothesis that Social Adaptability Index (SAI) based on employment, education, income, marital status and substance abuse is associated with survival in chronic kidney disease (CKD) patients. This is a retrospective cohort study of patients with CKD stage 2 or greater. We used the Third National Health and Nutrition Examination Survey (NHANES III) cohort data between 1988 and 1994 including those 18 years or older. Our primary variable of interest is SAI. Each component of SAI (employment status, education, marital status, and substance abuse) has been graded on the scale of 0-3, income has been graded on the scale 0-1. Age, sex, race, diabetes, co-morbidity index, body mass index (BMI), geographic location, haemoglobin, serum creatinine, serum albumin, serum cholesterol and Hba1c were used as covariates in multivariate analysis. The outcome of the study is patient's mortality. The time to death was calculated as time between the first interview by NHANES and death. We analysed 13 400 subjects with mean age of 50.6 ± 20-53.6% males, 44.4% white, 29.7% African American and 22.1% Mexican American-with 8.5% having diabetes, with an average number of co-morbid conditions of 2.7 ± 1.1. Lower SAI is associated with greater stage of CKD. Higher SAI was associated with decreased mortality [hazard ratio (HR) 0.88, P < 0.001, 95% confidence interval (CI) 0.86-0.89]. When SAI quintiles were analysed, we demonstrated a 'dose-dependent' association between SAI and survival. Subgroup analysis showed that this association of SAI and survival was present in all studied subgroups. The limitations of the study include retrospective design, potential misreporting and misclassification, and reverse causality. We demonstrated that SAI has a strong and clinically significant association with mortality in CKD patients.


Overcoming resistance to family-witnessed resuscitation.
Giving family members the option of being present during resuscitation has been shown to be beneficial for both family and staff. However, only a small percentage of intensive care units have policies promoting family-witnessed resuscitation. This article reviews current research showing the benefits of family-witnessed resuscitation, outlines how to successfully integrate a family facilitator during resuscitation, and includes research that has been effective in changing the prevailing attitudes of staff. The authors also argue for the resuscitation team to practice ethical and cultural humility when involving family members so that all resuscitation efforts are a success, whether the patient lives or dies.


2012 in review - part II: overcoming the obstacles in the pharma/biotech industry.
As highlighted in the first part of this review published last month, the year 2012 saw the approval of a remarkable number of new drugs, and among the new drugs reaching the market, a significant proportion were orphan drugs developed for treating less prevalent diseases. These drugs are certainly not expected to become blockbusters, but are of high interest because of their efficacy in a narrow spectrum of patients. This trend aligns with the general tendency of staying away from fit-for-all blockbusters into personalized medicine as one of the strategies for overcoming the patent cliff that resulted in a long list of drugs going off patent and being approved as generics also during last year. The emerging scenario resulting from new developments in the form of new drugs and biosimilars and newly available generic medications paralleled by strategic movements within the pharmaceutical industry to reinforce their position in the market, as reflected by merger and acquisition deals accompanied by significant efforts into prioritization resulting in spin-off and split transactions, is reviewed in this second part. This paper includes a significant amount of data in tables for quick review and to profile the new strategic movements in drug pipelines. Further information, including details on mechanisms of action, current status, itemized pharmacology, pharmacokinetic and clinical trial research findings and updated information can be found in the proprietary databases Thomson Reuters Integrity(SM) and Thomson Reuters Cortellis™.


Extraction of heavy metals by 2-mercaptoethoxy groups attached to silica gel.
Silica gel is currently being used as a support for various straight chain mono- and bifunctional compounds such as amines, thiols (mercaptans), and mercaptoalochols. The current study reports the results of an investigation of a 2-mercaptoethoxy moiety covalently attached to silica gel. The condensation was effected by acid-catalyzed condensation of toluene solution of 2-mercaptoethanol and silica gel and azeotropically removing the water using a Dean-Stark tube. As a coordinating agent, this bifunctional compound was used in the removal of copper(II), cadmium(II), and lead(II) from standard aqueous solutions. Using silica gel support, the removal percentages were 91-93% for the three metals.


The clinical significance of bilateral synostosis of the lambdoid suture and the usefulness of its treatment.
Extended craniectomy has been employed in 148 +/- 4 month old infants with bilateral synostosis of lambdoid suture. Intraoperatively the ICP was less than 180 mm H2O in 6, 180-200 in 1, greater than 200 in 7. 18 +/- 10 months following surgery clinical signs as psychomotor retardation and so forth were present in 4 compared to 11 patients preoperatively. With 1 exception there was a distinct remodelling of neurocranium and an almost complete ossification of the operative site.


Promoter Hypermethylation of GATA3, IL-4, and TGF-β Confers Susceptibility to Vogt-Koyanagi-Harada Disease in Han Chinese.
We investigated the role of promoter methylation of transcriptional and inflammatory factors, including TBX21, GATA3, RORγt, FOXP3, IFN-γ, IL-4, IL-17A, and TGF-β in the development of Vogt-Koyanagi-Harada (VKH) disease. The promoter methylation levels were detected by the Sequenom MassARRAY system in CD4+ T cells that were separated from 20 healthy individuals and 32 VKH patients (20 in the active stage without medication, 12 in inactive stage with medication). The mRNA expression level of GATA3, IL-4, and TGF-β in CD4+ T cells was analyzed by real-time RT-PCR. The promoter methylation levels of GATA3, IL-4, and TGF-β were significantly higher in active VKH patients than in healthy individuals (P < 0.05). A decreased mRNA expression of GATA3 and TGF-β was found in active VKH patients, which was correlated negatively with the DNA methylation of these factors. Treatment with systemic corticosteroid and cyclosporin A (CsA) decreased the methylation level of GATA3 and TGF-β in association with an increased mRNA expression of molecules and reduced disease activity. Our findings suggest that promoter hypermethylation of GATA3 and TGF-β in CD4+ T cells confers risk to VKH disease in Han Chinese.


The management of disseminated testicular cancer.
Fourteen patients with disseminated non-seminomatous germ cell tumours of the testis were treated with a 3-drug combination of vinblastine, bleomycin and cis-diamminedichloroplatinum. Of 13 patients with measurable disease, 9 (69%) had complete and 3 (23%) partial remission. One patient with a solitary metastasis failed to respond but is disease-free following surgical excision. Toxicity was significant but no drug-related deaths occurred and there were no apparent lasting side effects. No patient attaining complete remission has relapsed. Eleven (79%) of the 14 patients are without evidence of disease from 3+ to 36+ months, 6 of these for 2 years or more. Of the patients in partial remission one died at 16 months and one is undergoing further treatment for relapse with liver metastases. One patient remains in partial remission at 30 months. These results demonstrate the efficacy of this drug combination in testicular cancer.


Highly selective CO2 sensing response of lanthanum oxide nanoparticle electrodes at ambient temperature.
Lanthanum oxide nanoparticles (La2O3 NPs) are attractive rare earth metal oxides because of their applications in optical devices, catalysts, dielectric layers, and sensors. Herein, we report room temperature operative carbon dioxide gas sensing electrodes developed by a simple sonication assisted hydrothermal method. The physiochemical, morphological and gas-sensing properties of the prepared nanoparticles were studied systematically and their successful preparation was confirmed with the absence of impurities and high selectivity towards CO2. The fabricated sensor showed a high sensitivity of 40% towards CO2 at 50 ppm, and it can detect concentrations of up to 5 ppm with a quick response time of 6 s and recovery of 5 s. The electrode demonstrated long-term stability of 95% for 50 days when tested with an interval of 10 days. This simple and cost-effective method shows great potential for fabricating room temperature CO2 gas sensors.


Examination stress as an ecological inducer of cortisol and psychological responses to stress in undergraduate students.
The purpose of the present study was to investigate basic methodological issues related to the usage of an examination stress protocol in studies of psychoneuroendocrinology. In the present study, 57 undergraduate students served as participants. All subjects provided salivary samples and completed psychological inventories during a low examination stress period and again during a high examination stress period. Salivary samples were analyzed for cortisol. Three major findings were observed. First, the examination stress protocol proved to be an effective trigger of elevations in both psychological measures of stress and in cortisol levels. Second, sex differences were observed in cortisol levels, such that males showed an elevation in cortisol during the high examination stress session whereas females did not. Finally, no significant correlations were observed between elevations in psychological measures of stress and elevations in cortisol levels. These findings suggest that the examination stress protocol used in the present study effectively elevated both psychological stress and cortisol levels. Furthermore, these findings suggest that there are biological differences in how males and females respond to stress. Finally, no evidence was found to suggest a relationship between psychological and hormonal levels of stress. Together, these findings suggest the need to better define and consider the implications of both the specific measures of stress being used and individual differences in the subject samples in psychoendocrine studies.


Matching carotid anatomy with carotid stents.
Carotid artery stenting (CAS) has emerged as a promising minimal invasive treatment alternative to carotid arterectomy for patients with symptomatic and asymptomatic carotid artery stenosis. Complication rates of CAS continuously decreased during recent years. Operator experience and technical improvements equally contributed to the improved safety of the procedure. Today, devices specifically designed to fit the requirements of carotid interventions are available and allow a lesion-tailored interventional strategy. The choice of stents seems to play an important role in achieving optimal outcomes with the endovascular technique. Basically, three major types of stents are currently available: open cell nitinol stents, closed cell nitinol stents, and closed cell Elgiloy stents. The concept of cell design has implications on plaque coverage, but also on mechanical properties of the devices and conformation to carotid anatomy. The present review discusses the concept of lesion-tailored stenting to optimize early and late outcomes after CAS.


The role of ITPA and ribavirin transporter genes polymorphisms in prediction of ribavirin-induced anaemia in chronic hepatitis C Egyptian patients.
Few data are available concerning the roles of polymorphisms of inosine triphosphatase (ITPA) gene and ribavirin (RBV) transporter genes in the prediction of RBV-induced anaemia among Egyptians with chronic hepatitis C (CHC). Genotyping of three ITPA gene variants and two variants of RBV transporter genes has been performed in 123 patients under pegylated interferon-α/ribavirin treatment. The baseline haemoglobin and ITPA rs1127354 CA/AA have been found as predictors of anaemia at 4, 8 and 12 weeks of RBV therapy. In addition, ITPA rs7270101 AC/CC and age predicted anaemia after 12 weeks of therapy. In conclusion, the ITPA variant rs1127354C>A significantly predict RBV-induced anaemia during the first 3 months of treatment and it is recommended to be assessed before RBV administration.


Osseous changes of the mandibular condyle affect backward-rotation of the mandibular ramus in Angle Class II orthodontic patients with idiopathic condylar resorption of the temporomandibular joint.
Objective: To evaluate whether osseous changes of the temporomandibular joint (TMJ) condyle affect backward rotation of the mandibular ramus in Angle Class II orthodontic patients with idiopathic condylar resorption (ICR). Methods: Twenty Japanese women with Class II malocclusion with ICR (ICR group) and 24 women with Class II malocclusion without ICR (non-ICR group) were examined. Pre-treatment panoramic radiographs were used to measure condylar ratios. Pre-treatment lateral cephalograms were used to evaluate maxillofacial morphology. Results: The ICR group had a significantly smaller condylar ratio, greater backward rotation of the ramus, less labially inclined upper incisors, and a steeper occlusal plane. The increased backward rotation of the ramus in the ICR group was significantly associated with a smaller condylar ratio. Conclusion: Angle Class II patients with ICR had shorter condylar height attributable to osseous changes of the TMJ condyle, and the shorter condylar height may affect subsequent backward rotation of the ramus.


Aromatase inhibition for ovarian stimulation: future avenues for infertility management.
Ovarian stimulation is applied during infertility management either alone or in conjunction with intrauterine insemination and assisted reproductive technologies. At the present time, the two main medications used for ovarian stimulation include an oral antiestrogen, clomiphene citrate, and injectable gonadotropins. In spite of the high ovulation rate with the use of clomiphene citrate, the pregnancy rate is much lower. In clomiphene citrate failures, gonadotropin injections have generally been used as the next treatment option. Treatment with gonadotropins is difficult to control and characteristically associated with increased risk of severe ovarian hyperstimulation syndrome and high multiple pregnancies. Therefore, an effective oral treatment that could be used without risk of hyperstimulation and with minimal monitoring is the preferred therapy. We hypothesize that aromatase inhibitors can be administered early in the follicular phase to induce ovulation by releasing the hypothalamus or pituitary from estrogen negative feedback. Based on this hypothesis, we have reported the success of aromatase inhibitors in induction and augmentation of ovulation in addition to improving ovarian response to gonadotropin stimulation. Moreover, there are other potential applications for aromatase inhibitors in infertility management, including improving implantation in assisted reproduction and in-vitro maturation.


Oligoclonal banding in AIDS and hemophilia.
Hypergammaglobulinemia is a consistent finding in patients within the AIDS spectrum and with hemophilia A. Serum samples from patients with these conditions were analyzed for the presence of oligoclonal banding, using a high-resolution serum protein electrophoresis system. The incidence of banding is significantly greater in well homosexuals who are HIV-antibody positive and in patients with pre-AIDS-related complex, AIDS-related complex, AIDS with opportunistic infections, and AIDS with Kaposi's sarcoma than in normal blood donors. The incidence of banding is similar to controls in patients with hemophilia who have received either no blood products, cryoprecipitate only, or limited infusions of factor VIII concentrate. In patients who have received frequent infusions of factor VIII concentrate, the incidence of banding significantly increases. Thirteen of sixty-seven hemophiliac patients developed AIDS or symptoms related to HIV infection independent of their banding pattern. We hypothesize that the bands are not diagnostic of AIDS, but seem to correspond with disease progression, and that they are absent early in the disease, appear later in the course, and may disappear with advanced disease.


[A total removal case of cavernous angioma at the lateral wall of the third ventricle with interhemispheric trans-lamina terminalis approach].
A case of cavernous angioma at the lateral wall of the third ventricle which was totally removed with interhemispheric trans-lamina terminalis approach is reported. A 40-year-old male had a slowly progressive onset of partial diabetes insipidus and headache with no neurological deficit . CT scan revealed a high density area at anterior third ventricle. The tumor was diagnosed ectopic pinealoma because of CT findings and clinical symptoms. Irradiation and chemotherapy ( RAFP therapy) was performed to this lesion. After two months, his clinical symptoms disappeared. CT scan showed decrease of the density of the region at this point. He was discharged with no symptom. After a half year, he suddenly complained of right homonymous hemianopsia with headache. CT scan showed that the high density area became larger to left posterior than that of half year before. Left carotid angiogram showed no mass lesion and no abnormal vessel. Operation was performed with interhemispheric trans-lamina terminalis approach using bifrontal craniotomy. Operative findings revealed that the tumor situated at the lateral wall of the third ventricle, had rough surface with reddish colour, and old and fresh blood clots inside the tumor. The tumor was carefully dissected without brain damage and was totally removed. The histological findings was compatible with cavernous angioma. Post-operative CT scan showed no high density area. He was discharged with no neurological deficit without right homonymous hemianopsia. Cavernous angioma of anterior third ventricle is very rare.(ABSTRACT TRUNCATED AT 250 WORDS)


Synthesis and characterization of the core-shell magnetic molecularly imprinted polymers (Fe₃O₄@MIPs) adsorbents for effective extraction and determination of sulfonamides in the poultry feed.
In this study, we present a general method to prepare the core-shell magnetic molecularly imprinted polymers (MIPs) nanoparticles (NPs) for sulfamethazine (SMZ). The resulting Fe₃O₄@MIPs NPs possess a highly improved imprinting effect, fast adsorption kinetics and high adsorption capacity, and can be applied to extract sulfonamide in the poultry feed. In this protocol, the magnetite NPs were synthesized by co-precipitating Fe²⁺ and Fe³⁺ in an ammonia solution first. Silica was then coated on the Fe₃O₄ NPs using a sol-gel method to obtain silica shell magnetic NPs. Subsequently, the vinyl groups were grated onto silica-modified Fe₃O₄ surface by 3-methacryloyloxypropyltrimethoxysilane. Finally, the MIPs films were formed on the surface of Fe₃O₄@SiO₂ by the copolymerization of vinyl end groups with functional monomer, methacrylic acid, cross-linking agent, ethylene glycol dimethacrylate, the initiator azo-bis-isobutyronitrile and template molecule, sulfamethazine. The morphology, magnetic, adsorption and recognition properties of Fe₃O₄@MIPs NPs were characterized using transmission electron microscope (TEM), scanning electron microscope (SEM), Fourier transform infrared (FT-IR) spectrometer, vibrating sample magnetometer (VSM) and re-binding experiments. The results showed that the binding sites of Fe₃O₄@MIPs were good accessibility, fast adsorption rate and the maximum adsorption capacity of Fe₃O₄@MIPs to SMZ was 344.8 μg g⁻¹. The selectivity of the obtained Fe₃O₄@MIPs NPs were elucidated by the different rebinding capability of SMZ and structural related sulfonamides in the mixed solution. The results indicated that the Fe₃O₄@MIPs had high imprinting factor 9.5 and significant selectivity. A method was developed for enrichment and determination of SMZ in the poultry feed samples with recoveries of duck and chicken feed ranging from 63.3 to 76.5% and 68.7 to 74.7%, respectively and the relative standard deviations (RSD) (<6.7%).


Unilateral approach for bilateral clipping of posterior communicating artery aneurysms in a hybrid operating room: A technical note.
Bilateral posterior communicating (pComm) artery aneurysms represent only 2% of mirror intracranial aneurysms. Usually, these are surgically approached through bilateral craniotomies for clipping. We present the case of a 50-year-old female presenting with headache and horizontal diplopia. Neurological examination revealed a left oculomotor palsy, with no other neurological deficits. Imaging studies revealed bilateral aneurysmatic lesions in both internal carotid arteries (ICA). A conventional left pterional approach was planned in order to treat the symptomatic aneurysm, and, if deemed feasible, a contralateral clipping through the same approach. The procedure was performed in a hybrid operating room (HOR), performing an intraoperative digital subtraction angiography (DSA) and roadmapping assistance during dissection and clipping. Transoperatively, a post-fixed optic chiasm was identified, with a wide interoptic space, which allowed us to perform the contralateral clipping through a unilateral approach. This technique for clipping bilateral pComm aneurysms can be performed when the proper anatomical features are met.


Notch2 controls hepatocyte-derived cholangiocarcinoma formation in mice.
Liver cancer comprises a group of malignant tumors, among which hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC) are the most common. ICC is especially pernicious and associated with poor clinical outcome. Studies have shown that a subset of human ICCs may originate from mature hepatocytes. However, the mechanisms driving the trans-differentiation of hepatocytes into malignant cholangiocytes remain poorly defined. We adopted lineage tracing techniques and an established murine hepatocyte-derived ICC model by hydrodynamic injection of activated forms of AKT (myr-AKT) and Yap (YapS127A) proto-oncogenes. Wild-type, Notch1 flox/flox , and Notch2 flox/flox mice were used to investigate the role of canonical Notch signaling and Notch receptors in AKT/Yap-driven ICC formation. Human ICC and HCC cell lines were transfected with siRNA against Notch2 to determine whether Notch2 regulates biliary marker expression in liver tumor cells. We found that AKT/Yap-induced ICC formation is hepatocyte derived and this process is strictly dependent on the canonical Notch signaling pathway in vivo. Deletion of Notch2 in AKT/Yap-induced tumors switched the phenotype from ICC to hepatocellular adenoma-like lesions, while inactivation of Notch1 in hepatocytes did not result in significant histomorphological changes. Finally, in vitro studies revealed that Notch2 silencing in ICC and HCC cell lines down-regulates the expression of Sox9 and EpCAM biliary markers. Notch2 is the major determinant of hepatocyte-derived ICC formation in mice.


In-house work opportunities: implications for housing organizations serving persons living With HIV/AIDS.
Finding work and/or re-entering the workforce can be extremely challenging for persons living with HIV/AIDS. Also difficult is assisting them in the process, mostly because there is little documentation or resources about programs that provide vocational services specifically for this population. In response to this dilemma and because it was perceived as a win/win situation, three urban residential community organizations serving the HIV/AIDS population, decided independently to create in-house work opportunities for their clients. All of these organizations are a variation of the same theme: transitional/supportive housing for persons with HIV/AIDS that were formerly homeless and are now interested in becoming increasingly self-sufficient. This article will present a program description that addresses unique manner in which these three sites created in-house job programs in the areas of receptionist, kitchen, and maintenance work. More specifically, this paper will address the strengths, limitations, and ethical considerations that guided program development.


Phosphorylation and function of alpha4beta2 receptor.
The neuronal nicotinic acetylcholine receptor (nAChR) alpha4 and beta2 subunits expressed in heterologous expression systems assemble into high- and low-affinity receptors (Zwart and Vijverberg, 1998; Buisson and Bertrand, 2001; Houlihan et al., 2001; Nelson et al., 2003), which reflects the assembly of two distinct subunit stoichiometries of alpha4beta2 receptor (Nelson et al., 2003). The high-affinity receptor ([alpha4]2[beta2]3) is about 100-fold more sensitive to ACh than the low-affinity receptor ([alpha4]3[beta2]2) (Zwart and Vijverberg, 1998; Buisson and Bertrand, 2001; Houlihan et al., 2001; Nelson et al., 2003). Recent evidence implicated 14-3-3 proteins as modulators of the relative abundance of nAChR subunits in the endoplasmic reticulum (ER), where ligand-gated ion channels assemble. The 14-3-3 proteins influence ER-to-plasma membrane trafficking of multimeric cell-surface proteins (O'Kelly et al., 2002). 14-3-3 proteins bind components of these multimeric proteins, and this interaction overrides dibasic COP1 retention signal to permit forward transport of the protein (O'Kelly et al., 2002). In the case of alpha4beta2 nAChRs, 14-3-3 binds the alpha4 subunit, and this association is dependent on phosphorylation of a serine residue within a protein kinase A(PKA) consensus sequence in the large cytoplasmic domain of the alpha4 subunit, which is also a binding motif recognized by 14-3-3 (Jeancloss et al., 2001; O'Kelly et al., 2002). The interplay among PKA, alpha4 subunits, and 14-3-3 proteins increases cell-surface expression of alpha4beta2 nAChRs by increasing steady-state levels of the alpha4 subunit available for assembly with beta2 subunits (Jeancloss et al., 2001). Because it is not known how 14-3-3-dependent changes in the steady-state levels of the alpha4 subunit might affect the functional type of alpha4beta2 receptors, we have investigated the effects of mutations of the 14-3-3 binding motif in the alpha4 subunit on alpha4beta2 nAChR function.


Ultrasound Verification Of Safe Needle Examination Of The Rhomboid Major Muscle.
Palpation of a thoracic rib is a common method for reducing the risk of pneumothorax during electromyographic examination of the rhomboid major muscle, but its accuracy is unknown. Two physicians palpated healthy subjects to attempt to identify the center of a rib located beneath the rhomboid major muscle. The identified location was examined with ultrasonography to examine its accuracy and the subject's anatomical depths. Forty-four subjects (88 ribs) were studied. Palpation demonstrated a 66.3% accuracy rate, with significantly more incorrect palpations seen with greater muscle thickness (P = 0.004) and body mass index (P = 0.037), but not adipose thickness, age, or skin thickness (P > 0.05). Palpation of the ribs in an attempt to avoid inadvertent pneumothorax while examining the rhomboid major may be inaccurate, primarily in patients with large muscle bulk. We suggest a brief ultrasound evaluation before electromyography to gauge correct needle depth. Muscle Nerve 57: 61-64, 2018.


Lewis-base containing spiro type hole transporting materials for high-performance perovskite solar cells with efficiency approaching 20.
Owing to excellent performance and dopability, spiro-OMeTAD remains an irreplaceable hole transporting material (HTM) in perovskite solar cells (PSCs). In order to further improve the performance of spiro-OMeTAD based PSCs, a Lewis base can be introduced into the structure of spiro-OMeTAD wisely, which can keep the advantages of spiro-OMeTAD while incorporating the functionality of a Lewis base in passivating the surface of the perovskite. Therefore, spiro-type HTMs (spiro-CN-OMeTAD with a dicyano group and spiro-PS-OMeTAD with a thiocarbonyl group) were synthesized and confirmed by density functional theory (DFT) calculations and X-ray single-crystallographic diffraction. Spiro-CN-OMeTAD as an HTM is certified to have a suitable interfacial band alignment with the perovskite, good film quality and effective defect passivation, which facilitate the resulting device to achieve an efficiency of 19.90% with a high open-circuit voltage, low hysteresis, and improved stability. This study provides an alternative strategy for the molecular design of better HTMs in high-performance PSCs.


Aortoiliac reconstruction: the retroperitoneal approach and splenic injury.
Exposure of the abdominal aorta through the left retroperitoneum is a well-accepted technique. Unfortunately, splenic injury is a complication of this approach that rarely occurs through a mid-line transabdominal incision. In this series we evaluate the occurrence of splenic injury during left retroperitoneal aortic surgery. The records of all patients undergoing abdominal aortic reconstruction via the left retroperitoneal approach were reviewed from 1988 to 2001. Indications included either abdominal aortic aneurysm (AAA) or aortoiliac occlusive disease (AIOD). Thoracoabdominal aneurysms and visceral aortic reconstructions were excluded. Those patients that required splenectomy for splenic injury were stratified and analyzed for demographics, indications for operation, and other complications. Chi-square analysis was used to determine statistical significance. Over the 13-year study period, 2889 aortic reconstructions were performed. These consisted of 1773 elective AAA repairs, 357 ruptured or symptomatic AAA repairs, and 759 aortofemoral bypasses for AIOD. Splenectomies were performed after aortic repair in 21 (0.7%) patients. Breakdown by indication totaled 11/1773 (0.6%) for elective AAA repair, 7/357 (2.0%) for rupture or symptomatic AAA repair, and 3/759 (0.4%) for AIOD ( p < 0.05). Mortality for patients undergoing elective AAA without splenectomy was 1.9% (34/1762) and with splenectomy was 9.1% (1/11), while it was 3.2% (24/756) for AIOD patients without splenectomy and 14% (1/7) for AIOD patients with splenectomy (all p = NS). Splenic injury can occur in elective and emergent aortic reconstructions performed through the left retroperitoneum. The surgeon should be prepared for splenectomy whenever intraperitoneal blood or hemodynamic instability is identified. Evaluation of the spleen prior to flank closure may be prudent.


Perfusion magnetic resonance imaging of the liver.
Perfusion magnetic resonance imaging (MRI) studies quantify the microcirculatory status of liver parenchyma and liver lesions, and can be used for the detection of liver metastases, assessing the effectiveness of anti-angiogenic therapy, evaluating tumor viability after anti-cancer therapy or ablation, and diagnosis of liver cirrhosis and its severity. In this review, we discuss the basic concepts of perfusion MRI using tracer kinetic modeling, the common kinetic models applied for analyses, the MR scanning techniques, methods of data processing, and evidence that supports its use from published clinical and research studies. Technical standardization and further studies will help to establish and validate perfusion MRI as a clinical imaging modality.


High-purity enrichment of functional cardiovascular cells from human iPS cells.
A variety of human inherited heart diseases affect the normal functions of cardiomyocytes (CMs), endothelial cells (ECs), or smooth muscle cells (SMCs). To study human heart disease and generate cardiac cells for basic and translational research, an efficient strategy is needed for production of cardiac lineages from human stem cells. In the present study, a highly reproducible method was developed that can simultaneously enrich a large number of CMs and cardiac SMCs and ECs from human induced pluripotent stem (iPS) cells with high purity. Human multipotent cardiovascular progenitor cells were generated from human iPS cells, followed by selective differentiation of the multipotent cardiovascular progenitor cells into CMs, ECs, and SMCs. With further fluorescence-activated cell sorting, each of the three cardiovascular cell types could be enriched with high purity (>90%). These enriched cardiovascular cells exhibited specific gene expression signatures and normal functions when assayed both in vitro and in vivo. Moreover, CMs purified from iPS cells derived from a patient with LEOPARD syndrome, a disease characterized by cardiac hypertrophy, showed the expected up-regulated expression of genes associated with cardiac hypertrophy. Overall, our technical advance provides the means for generating a renewable resource of pure human cardiovascular cells that can be used to dissect the mechanisms of human inherited heart disease and for the future development of drug and cell therapeutics for heart disease.


Heat stress in pigs is accompanied by adipose tissue-specific responses that favor increased triglyceride storage.
Heat stress (HS) negatively affects all aspects of performance in pigs. Although certain tissue-specific responses in the liver, skeletal muscle, and intestine are known, there is paucity of information on responses within the adipose tissue. Therefore, the objective of this study was to delineate adipose tissue responses during HS in pigs. Thirty crossbred (Ossabaw × Duroc × Landrace) pigs were assigned to 3 treatments for 7 d. Treatments were 1) control and libitum fed (CON) with room temperature set at 20°C ± 1°C, 2) pair fed (PF) with room temperature as the CON treatment but pair fed to HS pigs, and 3) HS with room temperature 35°C ± 1°C and ad libitum access to feed. Compared with CON pigs, HS pigs had decreased feed intake and elevated skin temperature and respiration rate ( < 0.01). Blood urea nitrogen was higher ( = 0.01) in HS pigs compared with CON pigs only in males. In both subcutaneous and mesenteric adipose tissue, mRNA abundance of phosphoenolpyruvate carboxykinase (PCK1) was more elevated ( < 0.01) in HS groups compared with the CON and PF groups. Heat stress also caused increased heat shock protein 70 (HSP70; = 0.067) and CCAT/enhancer-binding homologous protein (CHOP) content ( < 0.05) in the mesenteric fat compared with the CON treatment. In conclusion, induction of PCK1 expression in adipose tissue by HS suggests elevated glyceroneogenesis might be involved in the increased fat storage in pigs under HS.


Reduced cartilage proteoglycan loss during zymosan-induced gonarthritis in NOS2-deficient mice and in anti-interleukin-1-treated wild-type mice with unabated joint inflammation.
To investigate the role of nitric oxide (NO) and interleukin-1 in (IL-1) joint inflammation and cartilage destruction during zymosan-induced gonarthritis (ZIA). Monarticular arthritis was elicited by intraarticular injection of zymosan. The effect of NO deficiency on arthritis was studied in mice with genetically disrupted NOS2. The role of IL-1 was examined by treating wild-type mice with neutralizing anti-murine IL-1(alpha+beta) antibodies. Joint swelling was measured externally by the increased uptake of circulating 99mtechnetium pertechnetate. Proteoglycan (PG) synthesis was assessed using 35S-sulfate incorporation into patellae ex vivo. Histology evaluated exudation and infiltration of leukocytes and the extent of cartilage destruction. The proinflammatory mediators NO, IL-1, and IL-6 were released by the articular tissues during the first hours of inflammation. Interestingly, anti-IL-1 treatment moderately reduced, and NOS2 deficiency moderately enhanced, joint swelling. However, the influx of neutrophils into the joint occurred independently of IL-1 and NOS2 activities. In the first week of inflammation, chondrocyte PG synthesis was significantly suppressed and chondrocytes became unresponsive to their essential anabolic factor, insulin-like growth factor 1 (IGF-1). Anti-IL-1 treatment or NOS2 deficiency prevented the inhibition of PG synthesis, and the chondrocytes remained IGF-1 responsive. Intraarticular injections of IL-1alpha into NOS2-deficient mice did not affect PG synthesis, thus proving that NO mediated this IL-1 effect in vivo. Furthermore, histology showed that cartilage PG loss was markedly ameliorated in NOS2-deficient and anti-IL-1-treated mice. Intermediate cartilage pathology was found in mice that were heterozygous for disrupted NOS2. IL-1 and NO play a minor role in edema and neutrophil influx, but a major role in cartilage destruction of ZIA. In this model of murine arthritis, cartilage destruction was, for the most part, caused by pronounced suppression of PG synthesis and IGF-1 unresponsiveness of the chondrocytes, which were induced by de novo-synthesized IL-1 and were mediated by NOS2 activation.


The tangential flow absorption model (TFAM) - A novel dissolution method for evaluating the performance of amorphous solid dispersions of poorly water-soluble actives.
There is a substantial demand for absorptive dissolution tests, as single vessel dissolution experiments were originally not designed for testing supersaturating systems. Current approaches suffer from inadequate mass transfer of the dissolved active from the dissolution site, discrepancies in the fluid volume compared to in vivo intestinal fluid volumes or the dilution of functional excipients. In this work a novel dissolution apparatus was developed that enables adjustable mass transfer of the active through a membrane, while retaining the functional polymeric excipients at the dissolution site. Using this setup the dissolution behavior of various spray dried amorphous solid dispersions containing carbamazepine, hydrochlorothiazide and ketoconazole as model actives at intermediate and high supersaturation levels was evaluated. Compared to non-absorptive dissolution experiments, differences in the concentration-time profiles were noted. The experiments with a high supersaturation of ketoconazole revealed a concentration decrease over time under absorptive conditions. Additionally, it was observed that the difference between "spring" as well as "spring and parachute" formulations was less pronounced with increasing drug efflux. Further, the apparatus was also tested with Fasted State Simulated Intestinal Fluid as dissolution medium and results were compared to phosphate buffer pH6.8. As major benefits of the new TFAM apparatus the easy experimental procedure and sample preparation for drug concentration measurements using spectroscopy in the permeate, without the necessity for additional filtration and/or centrifugation to remove precipitated drug molecules, could be highlighted. This TFAM approach seems to be a promising tool for identifying formulations for amorphous solid dispersions with optimal in vitro performance.


Safe dissection and complication avoidance for L1-2 interbody placement via a lateral access approach.
The lateral access approach for L1-2 interbody placement or other levels at or near the thoracolumbar junction may be difficult without proper knowledge and visualization of anatomy. Specifically, understanding where the fibers of the diaphragm travel and avoiding injury to the diaphragm are paramount. The video can be found here: https://stream.cadmore.media/r10.3171/2022.3.FOCVID2221.


Genetic aspects of erythropoietic protoporphyria.
An exhaustive study of erythropoietic protoporphyria in the Netherlands led to the discovery of 200 patients with this disorder in 91 families. In 46 of these families a single patient occurred. A study of parents, sibs and children led to the conclusion that the disease of erythropoietic protoporphyria is inherited as an autosomal recessive disorder. The presence of an occasional fluorescent red blood cell combined with normal protoporphyrin levels was observed in half of the children and sibs of patients and one of their parents; this phenomenon is therefore inherited as an autosomal dominant character. From an analysis of the findings in the 91 families we put forward the hypothesis of a 3-allele system.


Development and validation of an international risk prediction algorithm for episodes of major depression in general practice attendees: the PredictD study.
Strategies for prevention of depression are hindered by lack of evidence about the combined predictive effect of known risk factors. To develop a risk algorithm for onset of major depression. Cohort of adult general practice attendees followed up at 6 and 12 months. We measured 39 known risk factors to construct a risk model for onset of major depression using stepwise logistic regression. We corrected the model for overfitting and tested it in an external population. General practices in 6 European countries and in Chile. In Europe and Chile, 10 045 attendees were recruited April 2003 to February 2005. The algorithm was developed in 5216 European attendees who were not depressed at recruitment and had follow-up data on depression status. It was tested in 1732 patients in Chile who were not depressed at recruitment. Main Outcome Measure DSM-IV major depression. Sixty-six percent of people approached participated, of whom 89.5% participated again at 6 months and 85.9%, at 12 months. Nine of the 10 factors in the risk algorithm were age, sex, educational level achieved, results of lifetime screen for depression, family history of psychological difficulties, physical health and mental health subscale scores on the Short Form 12, unsupported difficulties in paid or unpaid work, and experiences of discrimination. Country was the tenth factor. The algorithm's average C index across countries was 0.790 (95% confidence interval [CI], 0.767-0.813). Effect size for difference in predicted log odds of depression between European attendees who became depressed and those who did not was 1.28 (95% CI, 1.17-1.40). Application of the algorithm in Chilean attendees resulted in a C index of 0.710 (95% CI, 0.670-0.749). This first risk algorithm for onset of major depression functions as well as similar risk algorithms for cardiovascular events and may be useful in prevention of depression.


Mortality and social class in Maori and nonMaori New Zealand men: changes between 1975-7 and 1985-7.
Social class mortality differences in Maori and nonMaori New Zealand men aged 15-64 years have previously been examined for the period 1975-7. The analysis has now been repeated for the period 1985-7 in order to examine changes over time. Numerator data were obtained from national death registrations and denominator data were obtained from the 1976 and 1986 censuses. These were used to calculate age-standardized death rates in Maori and nonMaori. Mortality declined by 28% in Maori and 14% in nonMaori between 1975-7 and 1985-7. The death rate for diseases amenable to medical intervention fell by 54% in Maori and 23% in nonMaori, but the Maori death rate was still 2.8 times the nonMaori death rate, whereas the relative risk for nonamenable causes was only 1.4. Some progress has been achieved in reducing ethnic differences in mortality in New Zealand men, but substantial differences remain for diseases which are amenable to medical intervention (including chronic rheumatic heart disease, hypertensive heart disease, and tuberculosis). It is likely that these differences reflect poor access to culturally safe and appropriate health care in Maori people.


Sp1 is a co-activator with Ets-1, and Net is an important repressor of the transcription of CTP:phosphocholine cytidylyltransferase alpha.
Phosphatidylcholine biosynthesis via the CDP-choline pathway is primarily regulated by CTP:phosphocholine cytidylyltransferase (CT) encoded by the Pcyt1a and Pcyt1b genes. Previously, we identified an Ets-1-binding site located at -49/-47 in the promoter of Pcyt1a as an important transcriptional element involved in basal CTalpha transcription (Sugimoto, H., Sugimoto, S., Tatei, K., Obinata, H., Bakovic, M., Izumi, T., and Vance, D. E. (2003) J. Biol. Chem. 278, 19716-19722). In this study, we determined whether or not there were other important elements and binding proteins for basal CTalpha transcription in the Pcyt1a promoter, and if other Ets family proteins bind to the Ets-1-binding site. The results indicate the formation of a ternary complex with Ets-1 binding at -49/-47 and Sp1 binding at -58/-54 of the Pcyt1a promoter that is important for activating CTalpha transcription. When nuclear extracts of COS-7 cells expressing various Ets family repressors were incubated with DNA probes, binding of Net to the probes was observed. Net dose-dependently depressed the promoter-luciferase activity by 98%, even when co-expressed with Ets-1. RNA interference targeting Net caused an increase of endogenous CTalpha mRNA. After synchronizing the cell cycle in NIH3T3 cells, CTalpha mRNA increased at the S-M phase corresponding to an increase of Ets-1 mRNA and a decrease of Net mRNA. These results indicated that Net is an important endogenous repressor for CTalpha transcription.


Adenocarcinoma in the lung in patients with breast cancer. A prospective analysis of the discriminatory value of immunohistology.
In some patients with a history of breast cancer who also have masses in the lung, making a clinical distinction between primary pulmonary neoplasia and pulmonary metastasis of mammary carcinoma may be impossible. To ascertain whether immunohistologic studies could contribute to resolving this problem, the authors undertook a prospective study of 30 cases showing synchronous or metachronous adenocarcinomas in these two sites. A predefined panel of antibodies--as derived from published antigenic catalogs for breast and lung cancer--was applied to each case. Tumors were interpreted as metastases if they were positive for gross cystic disease fluid protein-15, estrogen receptor protein, or S-100 protein. Conversely, primary adenocarcinomas of the lung were defined by their expression of carcinoembryonic antigen and a lack of the other three determinants. Using these criteria, 15 lesions were classified as metastatic; 11 were categorized as primary pulmonary adenocarcinomas; and 4 cases were indeterminate in origin. Responses to corresponding therapeutic protocols generally supported the validity of the immunohistologic diagnoses; 8 of 15 patients treated for metastatic breast cancer were well at least contact, as were 5 of 11 patients who received therapy for primary carcinoma of the lung. These data suggest that immunohistology plays a useful role in distinguishing mammary from pulmonary adenocarcinomas.


Characterization of the hom-thrC-thrB cluster in aminoethoxyvinylglycine-producing Streptomyces sp. NRRL 5331.
Three genes from the aminoethoxyvinylglycine (AVG)-producing Streptomyces sp. NRRL 5331 involved in threonine biosynthesis, hom, thrB and thrC, encoding homoserine dehydrogenase (HDH), homoserine kinase (HK) and threonine synthase (TS), respectively, have been cloned and sequenced. The hom and thrC genes appear to be organized in a bicistronic operon as deduced by disruption experiments. The thrB gene, however, is transcribed as a monocistronic transcript. The encoded proteins are quite similar to the HDH, HK and TS proteins from other bacterial species. The overall organization of these three genes, in the order hom-thrC-thrB, differs from that in other bacteria and is similar to that reported in the Streptomyces coelicolor genome sequence. This is the first time in which the gene cluster for the three last steps of threonine biosynthesis has been characterized from a streptomycete. Disruption of thrC indicated that threonine is not a direct precursor for AVG biosynthesis in Streptomyces sp. NRRL 5331 and suggested that the branching point of the aspartic acid-derived biosynthetic route of this metabolite should lie earlier on the threonine biosynthetic route.


Radioprotection of mice with interleukin-1: relationship to the number of spleen colony-forming units.
Compared to saline-injected mice 9 days after 6.5 Gy irradiation, there were twofold more Day 8 spleen colony-forming units (CFU-S) per femur and per spleen from B6D2F1 mice administered a radioprotective dose of human recombinant interleukin-1-alpha (rIL-1) 20 h prior to their irradiation. Studies in the present report compared the numbers of CFU-S in nonirradiated mice 20 h after saline or rIL-1 injection. Prior to irradiation, the number of Day 8 CFU-S was not significantly different in the bone marrow or spleens from saline-injected mice and rIL-1-injected mice. Also, in the bone marrow, the number of Day 12 CFU-S was similar for both groups of mice. Similar seeding efficiencies for CFU-S and percentage of CFU-S in S phase of the cell cycle provided further evidence that rIL-1 injection did not increase the number of CFU-S prior to irradiation. In a marrow repopulation assay, cellularity as well as the number of erythroid colony-forming units, erythroid burst-forming units, and granulocyte-macrophage colony-forming cells per femur of lethally irradiated mice were not increased in recipient mice of donor cells from rIL-1-injected mice. These results demonstrated that a twofold increase in the number of CFU-S at the time of irradiation was not necessary for the earlier recovery of CFU-S observed in mice irradiated with sublethal doses of radiation 20 h after rIL-1 injection.


Mass spectrometry-based characterization of the vitreous phosphoproteome.
the vitreous gel is a highly hydrated extracellular matrix containing many proteins. These proteins are likely accumulated in the vitreous by local secretion, filtration from the blood, or diffusion from the surrounding tissues and vasculature, and may be altered in disease state. In the last several years, several reports of large-scale profiling of vitreous proteins have been published; however, there is little information on the characterization of the phosphoproteome of vitreous. Here, we sought to identify phosphopeptides and their phosphorylation sites from vitreous. we used titanium dioxide (TiO(2) ) to enrich phosphopeptides from vitreous and identified them by LC-MS/MS. we identified 85 unique phosphopeptides and the phosphorylation sites from 44 proteins. we present a method for characterization of phosphoproteome from vitreous samples using current MS technologies and yielded an initial assessment of the phosphoprotein/peptide content of human vitreous, thus providing important biological information toward further understanding of the post-translational modifications of vitreous proteins and their functional significance in disease.


Constitutive IP3R1-mediated Ca2+ release reduces Ca2+ store content and stimulates mitochondrial metabolism in mouse GV oocytes.
In mammals, fertilization initiates Ca2+ oscillations in metaphase II oocytes, which are required for the activation of embryo development. Germinal vesicle (GV) oocytes also display Ca2+ oscillations, although these unfold spontaneously in the absence of any known agonist(s) and their function remains unclear. We found that the main intracellular store of Ca2+ in GV oocytes, the endoplasmic reticulum ([Ca2+]ER), constitutively 'leaks' Ca2+ through the type 1 inositol 1,4,5-trisphosphate receptor. The [Ca2+]ER leak ceases around the resumption of meiosis, the GV breakdown (GVBD) stage, which coincides with the first noticeable accumulation of Ca2+ in the stores. It also concurs with downregulation of the Ca2+ influx and termination of the oscillations, which seemed underpinned by the inactivation of the putative plasma membrane Ca2+ channels. Lastly, we demonstrate that mitochondria take up Ca2+ during the Ca2+ oscillations, mounting their own oscillations that stimulate the mitochondrial redox state and increase the ATP levels of GV oocytes. These distinct features of Ca2+ homeostasis in GV oocytes are likely to underpin the acquisition of both maturation and developmental competence, as well as fulfill stage-specific cellular functions during oocyte maturation.


Transforming growth factor β signaling overcomes dasatinib resistance in lung cancer.
Lung cancer is the second most common cancer and the leading cause of cancer-related deaths. Despite recent advances in the development of targeted therapies, patients with advanced disease remain incurable, mostly because metastatic non-small cell lung carcinomas (NSCLC) eventually become resistant to tyrosine kinase inhibitors (TKIs). Kinase inhibitors have the potential for target promiscuity because the kinase super family is the largest family of druggable genes that binds to a common substrate (ATP). As a result, TKIs often developed for a specific purpose have been found to act on other targets. Drug affinity chromatography has been used to show that dasatinib interacts with the TGFβ type I receptor (TβR-I), a serine-threonine kinase. To determine the potential biological relevance of this association, we studied the combined effects of dasatinib and TGFβ on lung cancer cell lines. We found that dasatinib treatment alone had very little effect; however, when NSCLC cell lines were treated with a combination of TGFβ and dasatinib, apoptosis was induced. Combined TGFβ-1 + dasatinib treatment had no effect on the activity of Smad2 or other non-canonical TGFβ intracellular mediators. Interestingly, combined TGFβ and dasatinib treatment resulted in a transient increase in p-Smad3 (seen after 3 hours). In addition, when NSCLC cells were treated with this combination, the pro-apoptotic protein BIM was up-regulated. Knockdown of the expression of Smad3 using Smad3 siRNA also resulted in a decrease in BIM protein, suggesting that TGFβ-1 + dasatinib-induced apoptosis is mediated by Smad3 regulation of BIM. Dasatinib is only effective in killing EGFR mutant cells, which is shown in only 10% of NSCLCs. Therefore, the observation that wild-type EGFR lung cancers can be manipulated to render them sensitive to killing by dasatinib could have important implications for devising innovative and potentially more efficacious treatment strategies for this disease.


Audit of transfusion among the oldest old: treading the fine line between undertransfusion and optimum trigger.
Recent research has questioned restrictive transfusion policies in vulnerable elderly populations. Our audit assesses the prevalence and postoperative outcomes of extremely elderly patients undergoing the stress of surgery with perioperative hemoglobin (Hb) less than 9 g/dL. This retrospective analysis of prospectively collected data addressed patients aged 85+ undergoing elective surgery. Demographic data and baseline characteristics were recorded, as well as Hb and transfused red blood cell (RBC) units. The main endpoint was the prevalence of perioperative Hb less than 9 g/dL, that is, patients with baseline Hb <9 g/dL without preoperative transfusions (defined as Group A). Patients with perioperative Hb of 9 g/dL or greater (with or without transfusion) were designated as Group B. Secondary outcomes included morbidity, length of hospital stay, and mortality 30 days and 6 months after surgery. A bivariate analysis was performed followed by logistic regression to determine whether undergoing the stress of surgery with perioperative Hb less than 9 g/dL was an independent risk factor for postoperative outcomes. A total of 148 patients were included. The prevalence of perioperative Hb less than 9 g/dL was 25%. It was associated with increased morbidity and mortality -both 30 days and 6 months after surgery- and a prolonged length of hospital stay. Anemia-associated complications were higher among patients from Group A, whereas transfusion-associated ones were evenly distributed. In all the regression models, perioperative Hb less than 9 g/dL was an independent risk factor for worse postoperative outcomes. Perioperative Hb less than 9 g/dL was common among patients aged 85+, and it was associated with increased risk of adverse postoperative outcomes. The tolerance to anemia might decrease perioperatively when Hb is less than 9 g/dL. Thus, less restrictive thresholds deserve further evaluation.


Cell separation and transportation between two miscible fluid streams using ultrasound.
This paper presents a two-stream microfluidic system for transporting cells or micro-sized particles from one fluid stream to another by acoustophoresis. The two fluid streams, one being the original suspension and the other being the destination fluid, flow parallel to each other in a microchannel. Using a half-wave acoustic standing wave across the channel width, cells or particles with positive acoustic contrast factors are moved to the destination fluid where the pressure nodal line lies. By controlling the relative flow rate of the two fluid streams, the pressure nodal line can be maintained at a specific offset from the fluid interface within the destination fluid. Using this transportation method, particles or cells of different sizes and mechanical properties can be separated. The cells experiencing a larger acoustic radiation force are separated and transported from the original suspension to the destination fluid stream. The other particles or cells experiencing a smaller acoustic radiation force continue flowing in the original solution. Experiments were conducted to demonstrate the effective separation of polystyrene microbeads of different sizes (3 μm and 10 μm) and waterborne parasites (Giardia lamblia and Cryptosporidium parvum). Diffusion occurs between the two miscible fluids, but it was found to have little effects on the transport and separation process, even when the two fluids have different density and speed of sound.


Celebrating 40 years of progress in bone marrow transplantation: a report from the 40th Annual Meeting of the European Society for Blood and Marrow Transplantation.
The European Society for Blood and Marrow Transplantation was established in 1974 to enable scientists and physicians involved in clinical bone marrow transplantation to share their experience and develop cooperative studies. The organization celebrated its 40th anniversary with a meeting that considered hematopoietic stem cell transplantation not as a standalone procedure, but as part of a complex therapeutic program managed by a multidisciplinary professional team. The role of antifungal prophylaxis, emerging resistance in Aspergillus, the management of mucormycosis and new guidelines on antifungal therapy were among the topics discussed by the physicians, nurses, allied health professionals and scientists attending the 40th Annual Meeting of the European Society for Blood and Marrow Transplantation.


Phage Diversity in the Human Gut Microbiome: a Taxonomist's Perspective.
Bacteriophages (phages) have been known for over a century, but only in the last 2 decades have we really come to appreciate how abundant and diverse they are. With that realization, research groups across the globe have shown the importance of phage-based processes in a myriad of environments, including the global oceans and soils, and as part of the human microbiome. Through advances in sequencing technology, genomics, and bioinformatics, we know that the morphological diversity of bacteriophages originally used for taxonomy is eclipsed by their genomic diversity. Because we currently do not have a complete taxonomic framework or naming scheme to describe this diversity, crucial information from virome and microbiome studies is being lost. In this commentary, I will discuss recent advances in taxonomy and its importance for studies of the microbiome with examples of the human gut phageome and make recommendations for future analyses.


Comparative Measurements of Radon Concentration in Soil Using Passive and Active Methods in High Level Natural Radiation Area (HLNRA) of Ramsar.
Radon and its daughters are amongst the most important sources of natural exposure in the world. Soil is one of the significant sources of radon/thoron due to both radium and thorium so that the emanated thoron from it may cause increased uncertainties in radon measurements. Recently, a diffusion chamber has been designed and optimized for passive discriminative measurements of radon/thoron concentrations in soil. In order to evaluate the capability of the passive method, some comparative measurements (with active methods) have been performed. The method is based upon measurements by a diffusion chamber, including two Lexan polycarbonate SSNTDs, which can discriminate the emanated radon/thorn from the soil by delay method. The comparative measurements have been done in ten selected points of HLNRA of Ramsar in Iran. The linear regression and correlation between the results of two methods have been studied. The results show that the radon concentrations are within the range of 12.1 to 165 kBq/m(3) values. The correlation between the results of active and passive methods was measured by 0.99 value. As well, the thoron concentrations have been measured between 1.9 to 29.5 kBq/m(3) values at the points. The sensitivity as well as the strong correlation with active measurements shows that the new low-cost passive method is appropriate for accurate seasonal measurements of radon and thoron concentration in soil.