Testing of In Vitro Susceptibility of Neisseria gonorrhoeae to Azithromycin: Comparison of Disk Diffusion and Reference Agar Dilution Methods.
Azithromycin in combination with ceftriaxone is recommended as the first-line treatment for uncomplicated gonorrhea in many countries. Therefore, monitoring of azithromycin susceptibility of Neisseria gonorrhoeae isolates is essential. In 2019, the Clinical and Laboratory Standards Institute (CLSI) listed the MIC breakpoint for a susceptible-only category to azithromycin, but breakpoints for disk diffusion are not yet available. In this study, we evaluated the usefulness of disk diffusion for testing the susceptibility of N. gonorrhoeae isolates to azithromycin. A total of 189 clinical isolates susceptible and nonsusceptible to azithromycin were used. Agar dilution MICs were correlated with inhibition zone diameters of azithromycin disks (15-μg) manufactured by BBL and Oxoid. In addition, an interlaboratory study involving two clinical microbiology laboratories was conducted. There was a strong correlation between disk diffusion and agar dilution for BBL disks (r = -0.74; P < 0.001) and Oxoid disks (r = -0.75; P < 0.001). Using a zone diameter breakpoint of ≥27 mm (susceptible) and ≤26 mm (nonsusceptible) yielded good separation between susceptible and nonsusceptible isolates and the least number of discrepancies. Compared to agar dilution, disk diffusion showed high agreement and kappa values of 95.2% and 0.899 (P < 0.001) for BBL disks and 96.8% and 0.933 (P < 0.001) for Oxoid disks, respectively. Major and very major discrepancies were observed in isolates with azithromycin MICs (1 and 2 μg/ml, respectively) near to the breakpoint. These data illustrate that disk diffusion could be a reliable method in clinical laboratories to test susceptibility to azithromycin in N. gonorrhoeae isolates.


CoMFA and CoMSIA analysis of tetrahydroquinolines as potential antimalarial agents.
Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were used on a dataset of compounds, some of them having been reported to inhibit Plasmodium falciparum protein, farnesyltransferase. The co-crystal structure of the lead molecule, BMS-214662 bound to Rat-PFT was used as a template. CoMFA yielded a good model, with r²(ncv) = 0.909, r²(cv) = 0.617 and was validated using an external set r²(pred) = 0.748). It compared favourably with CoMSIA. In the CoMFA model the steric and electrostatic fields exerted an almost equal influence on activity. The contour maps indicated the necessity for sterically large electropositive groups with electronegative tail to be present in these molecules for activity, and sterically large electronegative moieties on the sulfonamide linker. By incorporating these features some new compounds have been identified for further investigation.


Unveiling Contacts within Macromolecular Assemblies by Solving Minimum Weight Connectivity Inference (MWC) Problems.
Consider a set of oligomers listing the subunits involved in subcomplexes of a macromolecular assembly, obtained e.g. using native mass spectrometry or affinity purification. Given these oligomers, connectivity inference (CI) consists of finding the most plausible contacts between these subunits, and minimum connectivity inference (MCI) is the variant consisting of finding a set of contacts of smallest cardinality. MCI problems avoid speculating on the total number of contacts but yield a subset of all contacts and do not allow exploiting a priori information on the likelihood of individual contacts. In this context, we present two novel algorithms, MILP-W and MILP-WB. The former solves the minimum weight connectivity inference (MWCI), an optimization problem whose criterion mixes the number of contacts and their likelihood. The latter uses the former in a bootstrap fashion to improve the sensitivity and the specificity of solution sets.Experiments on three systems (yeast exosome, yeast proteasome lid, human eIF3), for which reference contacts are known (crystal structure, cryo electron microscopy, cross-linking), show that our algorithms predict contacts with high specificity and sensitivity, yielding a very significant improvement over previous work, typically a twofold increase in sensitivity.The software accompanying this paper is made available and should prove of ubiquitous interest whenever connectivity inference from oligomers is faced.


Intracorneal pustular drug eruption, a novel cutaneous adverse event in anti-programmed cell death-1 patients that highlights the effect of anti-programmed cell death-1 in neutrophils.
The introduction of anti-programmed cell death-1 (anti-PD1) monoclonal antibodies has revolutionized the treatment of various advanced malignancies. Despite its efficacy, anti-PD1 therapy is accompanied by a variety of cutaneous adverse events. A 79-year-old man developed erythematous scaly plaques and pustules of the forehead, legs and arms after four cycles of nivolumab infusions every 2 weeks. Histology showed intracorneal pustules with dermal neutrophils and eosinophils. He was treated successfully with topical corticosteroids without discontinuation of nivolumab. We report subcorneal pustular eruption as a novel cutaneous adverse event in patients on anti-PD1 therapy. Other neutrophilic eruptions (psoriasis, Sweet's syndrome, acute generalized pustulosis) have been reported in patients on anti-PD1 treatments, suggesting the neutrophil as another cell type modulated by anti-PD1 antibodies.


Adiabatic refocusing pulses for volume selection in magnetic resonance spectroscopic imaging.
Because of their excellent slice profiles and high immunity to RF inhomogeneity, adiabatic full passage (AFP) pulses are ideal for use in spatial localization. The nonlinear, position-dependent phase of a single AFP pulse generated during refocusing of transverse magnetization traditionally is eliminated by using identical pairs of AFP pulses, at the expense of increased RF power deposition and increased echo time (TE). Here it is shown that one can achieve significant phase refocusing by executing single AFP pulses along non-equivalent spatial axes. When used for volume selection in MR spectroscopic imaging (MRSI) the remaining nonlinear phase becomes inconsequential when the phase across a spectroscopic volume is small. Selection of rectangular and octagonal volumes is demonstrated with half the number of AFP pulses used in the traditional approach. It is shown that octagonal volume selection in the human brain provides excellent suppression of extracranial lipids, and thus allows multislice (1)H MRSI at 4 Tesla to be performed within the guidelines for RF power deposition.


Protective Pleiotropic Effect of Flavonoids on NAD⁺ Levels in Endothelial Cells Exposed to High Glucose.
NAD(+) is important for oxidative metabolism by serving as an electron transporter. Hyperglycemia decreases NAD(+) levels by activation of the polyol pathway and by overactivation of poly(ADP-ribose)-polymerase (PARP). We examined the protective role of three structurally related flavonoids (rutin, quercetin, and flavone) during high glucose conditions in an in vitro model using human umbilical vein endothelial cells (HUVECs). Additionally we assessed the ability of these flavonoids to inhibit aldose reductase enzyme activity. We have previously shown that flavonoids can inhibit PARP activation. Extending these studies, we here provide evidence that flavonoids are also able to protect endothelial cells against a high glucose induced decrease in NAD(+). In addition, we established that flavonoids are able to inhibit aldose reductase, the key enzyme in the polyol pathway. We conclude that this protective effect of flavonoids on NAD(+) levels is a combination of the flavonoids ability to inhibit both PARP activation and aldose reductase enzyme activity. This study shows that flavonoids, by a combination of effects, maintain the redox state of the cell during hyperglycemia. This mode of action enables flavonoids to ameliorate diabetic complications.


Traditional Chinese medicine syndrome distribution in chronic hepatitis B populations: a systematic review.
Many patients with chronic hepatitis B (CHB) seek help from traditional Chinese medicine (TCM). TCM treatment is based on syndrome differentiation. This study aimed to investigate the syndrome distribution in populations of CHB patients. The pre-specific search strategy was set, and 93 studies (20,106 participants) were identified by electronic and hand searches. The methodological quality of included studies was assessed. Data on syndrome distribution and correlations between syndromes and severity of CHB, were extracted and analyzed. Forty-seven syndromes were identified under 24 different syndrome diagnosis systems for CHB. The majority of included studies reported Liver Depression and Spleen Deficiency (LDSD) (52.54% of studies) or Liver-Gallbladder Dampness Heat (LGDH)/Dampness-Heat Obstructing Middle Energizer (DHME) (32.20%) as the major syndromes in CHB patients without cirrhosis. Pooled analysis revealed that LDSD and LGDH/DHME accounted for 61.19% of participants without cirrhosis. In addition, Liver-Kidney Yin Deficiency (LKYinD) (40.99%) and Spleen-Kidney Yang Deficiency (SKYangD) (40.43%) syndromes were common in patients with severe CHB. The percentage of patients with Blood Stasis syndrome increased as the disease progressed to cirrhosis (32.09%). To conclude, LDSD and LGDH/DHME syndromes are found in a significant majority of CHB patients, particularly in the early stages. LKYinD, SKYangD and Blood Stasis dominate in patients at more advanced stages. More epidemiological studies of high methodological quality on syndrome distribution in CHB and standardization of syndrome differentiation for CHB are required to confirm the trends indicated by the studies reviewed here; confirming these trends can increase the efficacy of treatment and give guidance to doctors.


Native valve infective endocarditis due to Achromobacter xylosoxidans in an apparently immunocompetent individual.
A 79-year-old woman presented with fever, lethargy and weight loss. Clinically, the patient was confused, frail and had a systolic murmur. Her temperature was 38 °C and she remained persistently febrile. Initial investigations revealed neutrophilia with an elevated C reactive protein level. Multiple peripheral blood cultures grew Achromobacter xylosoxidans, a Gram-negative rod, which is a very rare cause of infection in patients who are immunocompetent. Subsequent transoesophageal echocardiography confirmed endocarditis with obvious vegetations on the mitral valve. The patient was treated with intravenous meropenem and cotrimoxazole in line with microbiology guidance. Surgical intervention in the form of mitral valve replacement was considered, but the patient was felt to be at prohibitive risk. After 6 weeks of intravenous antibiotics, a repeat transoesophageal echocardiogram showed no improvement in the mitral valve vegetation, which had increased in size. At this stage, her clinical course was complicated by major upper gastrointestinal bleeding requiring transfusion, multiorgan failure and ultimately death.


[Damage to the transforming growth factor TGF-beta type II receptor gene and microsatellite instability in carcinoma cells of the gastrointestinal tract].
In order to compare the frequency of damage to the transforming growth factor TGF-beta receptor type II gene (RII gene) and microsatellite instability (MIN) in oncogenesis of sporadic and hereditary cancer of gastrointestinal tract (GIT), 4 groups of carcinomas were analyzed. They included sporadic gastric (GC), family gastric (FGC), sporadic colorectal (CC) and hereditary nonpolyposis colorectal (HNPCC) carcinomas having appropriate clinical and pathological characteristics. Each group consisted of two types of carcinomas, one of them showing MIN. The RII gene damage occurred in 89% of GC (8 cases out of 9), 86% of CC (6 out of 7), 71% of FGC (5 out of 7), 50% of HNPCC (3 out of 6) for carcinomas coupled with MIN, whereas only in 6% (1 out of 18) of GC and 5% (1 out of 22) of CC for carcinomas without MIN. No damage to RII gene was found in the cases of hereditary carcinomas which did not show MIN though the number of cases analyzed was not sufficient for final conclusions (3 cases of FGC and 3 HNPCC). The data revealed a correlation between the MIN phenotype and mutations in RII gene both for sporadic (p < 0.001) and for hereditary (p < 0.02) cases. For all 4 groups the frequency of RII gene damage was found for early and advanced carcinomas. This suggests that the deficiency of TGF-beta receptor complex in both sporadic and hereditary carcinomas of GIT is revealed at early stages of tumor development and consequently may be responsible for tumor progression. The correlation between RII gene damages and MIN in GIT carcinoma cells suggests that genetic change predetermined the neoplasia of colorectal and gastric epithelium and partially overlapped for both sporadic and hereditary cases.


Circumvention of methotrexate resistance in childhood leukemia subtypes by rationally designed antifolates.
Cellular methotrexate (MTX) resistance may cause treatment failure in childhood common/preB-acute lymphoblastic leukemia (c/preB-ALL), T-lineage ALL (T-ALL), and acute myeloid leukemia (AML). The ex vivo potency of several antifolates (MTX, trimetrexate [TMQ], GW1843U89, multitargeted antifolate [MTA], Raltitrexed, and ZD9331) was studied via in situ inhibition of thymidylate synthase (TS). After short-term exposure, relapsed c/preB-ALL (rALL, n = 21), T-ALL (n = 22), and AML (n = 22) were 3-fold, 10-fold, and 6-fold less sensitive to MTX (P </=.01) compared with initial c/preB-ALL (n = 43). This difference in resistance was not observed for TMQ. Also for GW1843U89 and MTA, no resistance was observed in rALL and AML compared with c/preB-ALL. T-ALL compared with c/preB-ALL tended to be less resistant to GW1843U89 (3-fold) and MTA (6-fold) than to MTX (10-fold) (P =.06). Raltitrexed was more active against c/preB-ALL compared with the other subtypes. After 21 hours continuous incubation, T-ALL and AML samples were equally sensitive as c/preB-ALL to MTX, but rALL was 3-fold resistant to MTX compared with initial c/preB-ALL (P =.003). The resistance of rALL was circumvented by TMQ (1-fold; P =.03) and GW1843U89 (1.4-fold; P =. 004). Novel antifolates, except MTA, displayed a more potent TS inhibition than MTX during continuous exposure. These results suggest that MTX resistance in AML and T-ALL can be circumvented by continuous exposure, and that novel antifolates should be explored further for MTX-resistant T-ALL, rALL, and AML cells.


Salivary duct carcinoma: correlation of morphologic features by fine needle aspiration cytology and histopathology.
Salivary duct carcinoma (SDC) is a highly aggressive primary salivary gland neoplasm that resembles intraductal and infiltrating breast carcinoma. To review cytomorphologic features of histology proven SDC and evaluate potential pitfalls in cytologic diagnosis. Fine needle aspiration cytology (FNAC) of five histologically proven SDCs were reviewed. One patient was an elderly male (61 years), while the other four patients were younger, in their fourth decade (average age: 38 years). The initial cytologic diagnoses in two of the cases were poorly differentiated carcinoma with differential diagnosis of SDC and high grade mucoepidermoid carcinoma, while in the third case, a possibility of malignant mixed tumor was suggested. In fourth and fifth cases, the diagnosis was suggestive of pleomorphic adenoma with cystic change. The spectrum of cytologic changes included flat sheets and cohesive papillary and three-dimensional clusters. There was moderate to severe nuclear pleomorphism and atypia. Cribriform pattern and necrosis were occasionally identified. Prominent bright granular metachromatic stroma was seen in two of the cases interpreted as pleomorphic adenoma with cystic change and in the tumor reported as suggestive of malignant mixed tumor. The fifth case showed numerous cyst macrophages and apocrine cells with mild nuclear atypia. FNAC of SDC is difficult to interpret because of overlapping cytomorphologic features. Bland cytomorphologic features in some cases and several clinical pitfalls are demonstrated in our series.


The microstructure of high--critical current superconducting films.
The microstructure of superconducting films that have shown high-critical current densities has been studied. The films are shown to be epitaxial and contain twins and precipitates. The main difference between these films and low current carrying samples is the absence of grain boundaries. These boundaries are therefore identified as the cause of the lower critical current in ceramic samples.


Prevotella histicola sp. nov., isolated from the human oral cavity.
Three strains of anaerobic, variably pigmenting, Gram-negative bacilli isolated from human oral mucosal tissue were subjected to a comprehensive range of phenotypic and genotypic tests and were found to comprise a homogeneous group. 16S rRNA gene sequence analysis and DNA-DNA hybridization revealed that the strains constituted a novel group within the genus Prevotella, being most closely related to Prevotella melaninogenica and Prevotella veroralis. A novel species, Prevotella histicola sp. nov., is proposed to accommodate these strains. Prevotella histicola is saccharolytic and produces acetic acid and succinic acid as major end products of fermentation and trace to minor amounts of isovaleric acid and lactic acid. The G+C content of the DNA of the type strain is 43 mol%. The type strain of Prevotella histicola is T05-04T (=DSM 19854T=CCUG 55407T).


HPV vaccination in a context of public mistrust and uncertainty: a systematic literature review of determinants of HPV vaccine hesitancy in Europe.
Europe is increasingly described as the region in the world with the least confidence in vaccination, and particularly in the safety of vaccines. The aim of this systematic literature review was to gather and summarise all peer-reviewed and grey literature published about determinants of Human Papillomavirus (HPV) vaccine hesitancy in Europe. Ten thematic categories were identified across the 103 articles which were included in the review. Participants from European studies most commonly reported issues with the quantity and quality of information available about HPV vaccination; followed by concerns about potential side effects of the vaccine; and mistrust of health authorities, healthcare workers, and new vaccines. Comparative analyses indicated that confidence determinants differed by country and population groups. This evidence supports the need to develop context-specific interventions to improve confidence in HPV vaccination and design community engagement strategies aiming to build public trust.


Relation between phase of menstrual cycle and emergency department visits for acute asthma.
Hormonal fluctuations during the menstrual cycle are hypothesized to influence the course of asthma among women. A recent study found that almost 50% of emergency department (ED) visits occur during the perimenstrual phase. Our prospective cohort study in 64 EDs examined the relation between phase of menstrual cycle and visits for acute asthma. A total of 288 women with acute asthma were evaluated with a standardized patient interview and medical record review after excluding subjects who were pregnant, on hormonal therapy, postmenopausal, status post hysterectomy, had incomplete reproductive data, or whose ED visit fell more than 28 d after their last menstrual period. Only 13% reported reproductive factors as a personal asthma trigger. For all subjects, ED asthma visits were classified by menstrual phase: 33% were preovulatory (Days 5 to 11), 26% were periovulatory (Days 12 to 18), 20% were postovulatory (Days 19 to 25), and 21% were perimenstrual (Days 26 to 4), p = 0.008. There was no significant association between phase of menstrual cycle and asthma severity. Our data indicate that ED visits for acute asthma among women are more frequent during the preovulatory phase in contrast to other studies reporting more visits in the perimenstrual phase.


Association of fluid overload with kidney disease progression in advanced CKD: a prospective cohort study.
Fluid overload is a common phenomenon in patients in a late stage of chronic kidney disease (CKD). However, little is known about whether fluid overload is related to kidney disease progression in patients with CKD. Accordingly, the aim of the study was to assess the association of the severity of fluid status and kidney disease progression in an advanced CKD cohort. Prospective observational cohort study. This cohort study enrolled 472 non-dialysis-dependent patients with CKD stages 4-5 who were in an integrated CKD care program from January 2011 to December 2011 and followed up until December 2012 or initiation of renal replacement therapy (RRT). Tertile of fluid overload, with cutoff values at 0.6 and 1.6 L. RRT, rapid estimated glomerular filtration rate (eGFR) decline (faster than 3 mL/min/1.73 m(2) per year), and change in eGFR. The severity of fluid overload was measured by a bioimpedance spectroscopy method. eGFR was computed using the 4-variable MDRD (Modification of Diet in Renal Disease) Study equation. During a median 17.3-month follow-up, 71 (15.0%) patients initiated RRT and 187 (39.6%) experienced rapid eGFR decline. The severity of fluid overload was associated with increased risk of RRT (tertile 3 vs tertile 1: adjusted HR, 3.16 [95% CI, 1.33-7.50]). Fluid overload value was associated with increased risk of rapid eGFR decline (tertile 3 vs tertile 1: adjusted OR, 4.68 [95% CI, 2.30-9.52]). Furthermore, the linear mixed-effects model showed that the reduction in eGFR over time was faster in tertile 3 than in tertile 1 (P=0.02). The effect of fluid volume variation over time must be considered. Fluid overload is an independent risk factor associated with initiation of RRT and rapid eGFR decline in patients with advanced CKD.


Mutations in 12 known dominant disease-causing genes clarify many congenital anomalies of the kidney and urinary tract.
Congenital anomalies of the kidney and urinary tract (CAKUT) account for approximately half of children with chronic kidney disease. CAKUT can be caused by monogenic mutations; however, data are lacking on their frequency. Genetic diagnosis has been hampered by genetic heterogeneity and lack of genotype-phenotype correlation. To determine the percentage of cases with CAKUT that can be explained by mutations in known CAKUT genes, we analyzed the coding exons of the 17 known dominant CAKUT-causing genes in a cohort of 749 individuals from 650 families with CAKUT. The most common phenotypes in this CAKUT cohort were vesicoureteral reflux in 288 patients, renal hypodysplasia in 120 patients, and unilateral renal agenesis in 90 patients. We identified 37 different heterozygous mutations (33 novel) in 12 of the 17 known genes in 47 patients from 41 of the 650 families (6.3%). These mutations include (number of families): BMP7 (1), CDC5L (1), CHD1L (5), EYA1 (3), GATA3 (2), HNF1B (6), PAX2 (5), RET (3), ROBO2 (4), SALL1 (9), SIX2 (1), and SIX5 (1). Furthermore, several mutations previously reported to be disease-causing are most likely benign variants. Thus, in a large cohort over 6% of families with isolated CAKUT are caused by a mutation in 12 of 17 dominant CAKUT genes. Our report represents one of the most in-depth diagnostic studies of monogenic causes of isolated CAKUT in children.


Implementation of evidence-based practices for surgical site infection prophylaxis: results of a pre- and postintervention study.
Although evidence-based guidelines for best practices pertaining to surgical site infection (SSI) prophylaxis exist, the feasibility of implementing such practices remains to be demonstrated outside of a controlled clinical trial. This study was designed to assess the safety and feasibility of implementing evidence-based care practices to prevent SSIs. A prospective, double-cohort (pre- and postintervention) trial in elective, general surgery patients was conducted. All patients undergoing elective, major colorectal or hepatobiliary operations were enrolled. Postintervention cohort patients were exposed to new strategies to improve antibiotic administration times, perioperative normothermia rates, and perioperative glucose control. They were compared with the preintervention cohort, which received standard practice at the time. Outcomes evaluated include timing of antibiotic administration, perioperative temperatures, and postoperative glucose levels. SSI rates between cohorts were also compared. A total of 208 patients were enrolled. The proportion of patients receiving their preoperative antibiotics within 60 minutes improved from 5.9% to 92.6% (p < 0.001); perioperative normothermia rates improved from 60.5% to 97.6% (p < 0.001) between cohorts. There was no improvement in rates of hyperglycemia. SSI rates improved but did not reach statistical significance (14.3% versus 8.7%; p = 0.21). Implementation of evidence-based care practices to prevent SSI is both safe and practical outside the setting of a randomized, controlled trial. Sustained compliance remains to be demonstrated, although practice audits at our institution suggest ongoing success is possible.


Compliance with the Health Information and Quality Authority of Ireland National Standard for Patient Discharge Summary Information: a retrospective study in secondary care.
Unexplained changes to medication are common at hospital discharge and underscore the need to standardise patient discharge clinical documentation. In 2013, the Health Information and Quality Authority in Ireland published a Standard on the structure and content of discharge summaries. The intention was to ensure that all necessary information was complete and communicated to the next care provider. This study investigated one Hospital's compliance with the Standard, and appraised two methods of electronic discharge communication (Symphony or Tallaght Education and Audit Management System (TEAMS)). A retrospective survey of 198 randomly selected discharge summaries was conducted at the study hospital, a 600 bed academic teaching hospital located in Dublin, Ireland. Of the 198 evaluated summaries, mean total compliance was 77%±4.2 (95% CI 76.3 to 77.5). Most (84.7%, n=173) summaries were completed using one of the systems (TEAMS). Absence of communication about alteration of preadmission medication was frequent (107 out of 130 patients (82.3%, CI 76.2 to 89.2)). Higher compliance rates were observed however, when information was interfaced or where there were dedicated fields to be completed. Efforts to improve compliance with the National Standard for Patient Discharge Summary Information should focus on reporting changes made to medication during hospitalisation.


Carotenoids in floral parts of a narcissus, a daffodil and a tulip.
1. The qualitative and quantitative distribution of carotenoids of the floral parts of three monocotyledons, the narcissus Scarlet Elegance, the daffodil King Alfred and the tulip Golden Harvest, were studied. beta-Carotene, lutein or epoxy-beta-carotenes were usually the main pigments, depending on the floral part and on the flower. When beta-carotene was the major pigment there were only small amounts of, or sometimes no, epoxycarotenes. 2. Anthers, stigmas and styles of the three flowers did not possess any specific carotenoids but in some cases contained appreciable amounts of epoxycarotenoids. The possibility that these take part in reproduction is discussed. 3. The generalization that yellow flowers contained large amounts of xanthophylls and only traces of carotenes, whereas deep-orange flowers seemed to be characterized by the presence of large amounts of one carotene, was not always the correct one. It is suggested that the floral parts are yellow or orange depending on what carotenoids are present, which is the major one and the amount of total carotenoids, and also on the presence of other non-carotenoid pigments. 4. Two new probable isomers of 5,6:5',6'-diepoxy-beta-carotene were isolated and found together in various floral parts of the tulip Golden Harvest.


[Nutritional status in uremic patients treated with continuous ambulatory peritoneal dialysis. Relation with 1 year's evolution].
Signs of malnutrition have been detected in patients with chronic renal failure and a possible relation with complications of the disease has been speculated. The aim of the present study was to analyze the nutritional status of a group of patients with the aforementioned diagnosis on a substitutive treatment program with outpatient continuous peritoneal dialysis (CAPD) and its relation with complications over one year. The nutritional status of 29 of these patients and 22 individuals with normal renal function (controls) with global parameters of body fat, muscular protein and visceral protein being evaluated. The study was repeated at 6 and 12 months after the initial evaluation. The incidence of mortality, need for hospital admission because of complication and episodes of clinical peritonitis were analyzed. More than 70% of the patients presented protein-caloric malnutrition to a greater or lesser degree. This percentage dit not modify throughout the study and was produced at the expense of visceral protein, additionally affecting the fat and muscular protein of the males. Mortality, need for admission and the number of cases of peritonitis were greater in those presenting the lowest fat parameters with these complications beings foreseen from determined fat values. These patients have malnutrition which is related with the complications in the course of substitutive therapy making surveillance and nutritional support necessary.


Acquired Resistance to FGFR Inhibitor in Diffuse-Type Gastric Cancer through an AKT-Independent PKC-Mediated Phosphorylation of GSK3β.
Preclinical models of diffuse-type gastric cancer (DGC) that reliably predict clinical activity of novel compounds are lacking. To overcome the problem of poor tumor cellularity in DGC, we used cells from malignant ascites to establish DGC patient-derived xenograft (PDX) models that recapitulate the primary cancer. Cells in PDX model GAGA6 with FGFR2 amplification were sensitive to AZD4547, a potent FGFR inhibitor that is being clinically evaluated for FGFR-aberrant cancer types. Intermittent in vivo treatment of GAGA6 tumors with AZD4547 gave rise to PDX tumors with acquired resistance to AZD4547, GAGA6-R. Surprisingly, there were no mutations in the FGFR2 gene in GAGA6-R, negating gatekeeper mutations as a mechanism of drug resistance. Phosphorylation of FGFR2 and downstream signaling molecules AKT/PKB and MAPK/ERK remained inhibited by AZD4547. Further analysis of signaling pathways identified AKT-independent phosphorylation and inhibition of GSK3β as a mechanism of drug resistance in GAGA6-R cells. Treatment of GAGA6-R cells with protein kinase C (PKC) inhibitor H7 in combination with AZD4547 led to dephosphorylation and activation of GSK3β with concomitant downregulation of MCL-1 and BCL-XL. Combined treatment with AZD4547 and H7 in vitro synergistically enhanced cell death in GAGA6-R but not GAGA6 cells. Furthermore, midostaurin, a multikinase inhibitor with PKC-inhibiting activity, in part reversed resistance of GAGA6-R tumor to AZD4547 in vivo Our results suggest that upon challenge with FGFR inhibitors, FGFR2-amplified tumors that are highly dependent on FGFR2 signaling for survival rapidly develop resistance by switching to a PKC-mediated inhibition of GSK3β to gain a survival advantage. Mol Cancer Ther; 17(1); 232-42. ©2017 AACR.


Uncleaved ApoM signal peptide is required for formation of large ApoM/sphingosine 1-phosphate (S1P)-enriched HDL particles.
Apolipoprotein M (apoM), a plasma sphingosine 1-phosphate (S1P) carrier, associates with plasma HDL via its uncleaved signal peptide. Hepatocyte-specific apoM overexpression in mice stimulates formation of both larger nascent HDL in hepatocytes and larger mature apoM/S1P-enriched HDL particles in plasma by enhancing hepatic S1P synthesis and secretion. Mutagenesis of apoM glutamine 22 to alanine (apoM(Q22A)) introduces a functional signal peptidase cleavage site. Expression of apoM(Q22A) in ABCA1-expressing HEK293 cells resulted in the formation of smaller nascent HDL particles compared with wild type apoM (apoM(WT)). When apoM(Q22A) was expressed in vivo, using recombinant adenoviruses, smaller plasma HDL particles and decreased plasma S1P and apoM were observed relative to expression of apoM(WT). Hepatocytes isolated from both apoM(WT)- and apoM(Q22A)-expressing mice displayed an equivalent increase in cellular levels of S1P, relative to LacZ controls; however, relative to apoM(WT), apoM(Q22A) hepatocytes displayed more rapid apoM and S1P secretion but minimal apoM(Q22A) bound to nascent lipoproteins. Pharmacologic inhibition of ceramide synthesis increased cellular sphingosine and S1P but not medium S1P in both apoM(WT) and apoM(Q22A) hepatocytes. We conclude that apoM secretion is rate-limiting for hepatocyte S1P secretion and that its uncleaved signal peptide delays apoM trafficking out of the cell, promoting formation of larger nascent apoM- and S1P-enriched HDL particles that are probably precursors of larger apoM/S1P-enriched plasma HDL.


Study of LH response to GnRH in the young male as a criterion of genetic merit for female reproduction in sheep.
A high and a low response line in sheep were selected on the basis of the mean concentration of LH in 10-week-old Finn-Dorset ram lambs after an i.v. injection of 5 micrograms GnRH. After 8 male generations the mean LH response of the high line was more than 5-fold that of the low line and the heritability of the selected trait was estimated at 0.44 +/- 0.015. Highly significant line differences in mean LH response to GnRH were also found in males at 20 weeks of age and females at 10 and 20 weeks of age and the genetic correlations between the four LH response traits appear to be close to unity. Large line differences in the mean FSH response to GnRH were also found in both males and females at 10 and 20 weeks of age. Selection had little effect on the physical characteristics of lambs. High-response line ewes entering their first breeding season at about 7 months of age showed oestrus earlier in the season and had higher ovulation rates and numbers of lambs born per ewe lambing than did low-response line ewes. In the second breeding season, at about 19 months of age, the only line difference was a higher ovulation rate early in the breeding season in high-line ewes. It is suggested that these changes may be mediated by a more rapid response in high-line ewes to increased GnRH stimulation at puberty or at the beginning of the breeding season.


Leaf nodule endosymbiotic Burkholderia confer targeted allelopathy to their Psychotria hosts.
After a century of investigations, the function of the obligate betaproteobacterial endosymbionts accommodated in leaf nodules of tropical Rubiaceae remained enigmatic. We report that the α-D-glucose analogue (+)-streptol, systemically supplied by mature Ca. Burkholderia kirkii nodules to their Psychotria hosts, exhibits potent and selective root growth inhibiting activity. We provide compelling evidence that (+)-streptol specifically affects meristematic root cells transitioning to anisotropic elongation by disrupting cell wall organization in a mechanism of action that is distinct from canonical cellulose biosynthesis inhibitors. We observed no inhibitory or cytotoxic effects on organisms other than seed plants, further suggesting (+)-streptol as a bona fide allelochemical. We propose that the suppression of growth of plant competitors is a major driver of the formation and maintenance of the Psychotria-Burkholderia association. In addition to potential agricultural applications as a herbicidal agent, (+)-streptol might also prove useful to dissect plant cell and organ growth processes.


Surface-Immobilized ZnNx Sites as High-Performance Catalysts for Continuous Flow Knoevenagel Condensation in Water.
By immobilizing the metal complex on the substrate surface, our previous results have demonstrated that heterogeneous catalysts with well-dispersed active MNC (metal-nitrogen-carbon) sites can be prepared in a rational and efficient manner. In this study, we employed agarose aerogel (AA) as the substrate to illustrate a straightforward strategy for immobilizing ZnNx sites on the surface. Under relatively low temperatures, the amine group of the ligand condenses with the surface carbonyl group generated in situ, resulting in the surface immobilized Zn sites. This can be supported by the IR, PXRD, and XPS data. Comprehensive characterization methods, including synchrotron powder XRD and spherical aberration-corrected TEM, confirmed the absence of ZnNx site aggregation in the surface immobilization process, even with a high Zn content (up to 8 wt %). The immobilized ZnNx sites exhibited high catalytic performance in Knoevenagel condensation, and α,β-unsaturated compounds were obtained with high yield in both batch and continuous flow reactions. AA-ZnNx-200 showed the best catalytic activity, which was processed under 200 °C with a Zn content of 4.62 wt %. The immobilized ZnNx sites activated both the aldehyde and nitrile substrates, which were quantitatively converted into the corresponding α,β-unsaturated compounds, with water as the solvent at room temperature. In continuous flow reaction conditions, a conversion rate up to 99% can be achieved with malononitrile. This heterogeneous catalyst can be facilely produced with quantitative yield in a large scale from cheap starting material under mild conditions. No catalyst deactivation was observed after seven batch reaction cycles or 80 h of continuous flow reaction, indicating its high robustness under catalytic reaction conditions. This catalyst enables a separation-free, energy-saving, and environment-friendly production process, offering a practical way for the industrial production.


Advantages and limitations of genomics in prokaryotic taxonomy.
Taxonomic classification is an important field of microbiology, as it enables scientists to identify prokaryotes worldwide. Although the current classification system is still based on the one designed by Carolus Linnaeus, the currently available genomic content of several thousands of sequenced prokaryotic genomes represents a unique source of taxonomic information that should not be ignored. In addition, the development of faster, cheaper and improved sequencing methods has made genomics a tool that has a place in the workflow of a routine microbiology laboratory. Thus, genomics has reached a stage where it may be used in prokaryotic taxonomic classification, with criteria such as the genome index of average nucleotide identity being an alternative to DNA-DNA hybridization. However, several hurdles remain, including the lack of genomic sequences of many prokaryotic taxonomic representatives, and consensus procedures to describe new prokaryotic taxa that do not, as yet, accommodate genomic data. We herein review the advantages and disadvantages of using genomics in prokaryotic taxonomy.


Neuropsychological impairments in schizophrenia and psychotic bipolar disorder: findings from the Bipolar-Schizophrenia Network on Intermediate Phenotypes (B-SNIP) study.
Familial neuropsychological deficits are well established in schizophrenia but remain less well characterized in other psychotic disorders. This study from the Bipolar-Schizophrenia Network on Intermediate Phenotypes (B-SNIP) consortium 1) compares cognitive impairment in schizophrenia and bipolar disorder with psychosis, 2) tests a continuum model of cognitive dysfunction in psychotic disorders, 3) reports familiality of cognitive impairments across psychotic disorders, and 4) evaluates cognitive impairment among nonpsychotic relatives with and without cluster A personality traits. Participants included probands with schizophrenia (N=293), psychotic bipolar disorder (N=227), schizoaffective disorder (manic, N=110; depressed, N=55), their first-degree relatives (N=316, N=259, N=133, and N=64, respectively), and healthy comparison subjects (N=295). All participants completed the Brief Assessment of Cognition in Schizophrenia (BACS) neuropsychological battery. Cognitive impairments among psychotic probands, compared to healthy comparison subjects, were progressively greater from bipolar disorder (z=-0.77) to schizoaffective disorder (manic z=-1.08; depressed z=-1.25) to schizophrenia (z=-1.42). Profiles across subtests of the BACS were similar across disorders. Familiality of deficits was significant and comparable in schizophrenia and bipolar disorder. Of particular interest were similar levels of neuropsychological deficits in relatives with elevated cluster A personality traits across proband diagnoses. Nonpsychotic relatives of schizophrenia probands without these personality traits exhibited significant cognitive impairments, while relatives of bipolar probands did not. Robust cognitive deficits are present and familial in schizophrenia and psychotic bipolar disorder. Severity of cognitive impairments across psychotic disorders was consistent with a continuum model, in which more prominent affective features and less enduring psychosis were associated with less cognitive impairment. Cognitive dysfunction in first-degree relatives is more closely related to psychosis-spectrum personality disorder traits in psychotic bipolar disorder than in schizophrenia.


A Year After Implementation of the Telehealth Waiver: Being Offered and Utilizing Video-Specific Telehealth Among Dual-Eligible Medicare Recipients During the COVID-19 Pandemic.
Telehealth is an essential tool to provide access to care while reducing infection exposure for high-risk populations during the COVID-19 pandemic. Our study aims to examine factors associated with telehealth availability and usage among Medicare and dual-eligible recipients 1 year after implementation of the Medicare's temporary telehealth waiver. A cross-sectional, phone survey with a national representative sample of Medicare recipients. We obtained a final study sample from the Winter 2021 COVID-19 Supplement of Medicare Current Beneficiary Survey dataset (N = 10 586). We examined associations for being offered and having had telehealth visits or any video telehealth visits during the pandemic since November 1, 2020. Our primary outcomes were being offered any telehealth, being offered any video telehealth, having had any telehealth visit, and having had any video telehealth. Although dual eligibility was not significantly associated with being offered or having had any telehealth services during the pandemic, those who were dual eligible were more likely to have had video telehealth visits (adjusted odds ratio = 1.39, 95% confidence interval 1.04-1.86, P = .03) compared with those with non-dual eligibility. Recipients with disability eligibility, technology access, and severe chronic conditions were more likely to have been offered or have had telehealth. At the same time, those who lived in the nonmetropolitan area were less likely to have been offered or have had telehealth, including video telehealth. Our findings suggest that the federal waivers to expand telehealth services were successful in continuing care for vulnerable Medicare recipients. The providers' specific outreach and intervention efforts to offer telehealth visits are crucial for dual-eligible recipients. To increase video telehealth uptake, technology access and services to rural areas should be prioritized.


An Exploration of Tri-Axial Accelerometers in Assessing the Therapeutic Efficacy of Constraint-Induced Movement Therapy in Children with Unilateral Cerebral Palsy.
This study aimed to elucidate the role of tri-axial accelerometers in assessing therapeutic interventions, specifically constraint-induced movement therapy (CIMT), in children with unilateral cerebral palsy (UCP). The primary focus was understanding the correlation between the actigraphy metrics recorded during CIMT sessions and the resultant therapeutic outcomes. Children with UCP, aged between 4 and 12 years, participated in this study from July 2021 to December 2022. In conjunction with in-clinic sessions, during which participants wore tri-axial accelerometers on both limbs, we analyzed actigraphy data over three days of routine activities pre- and post-CIMT. While not all metrics derived from the accelerometers indicated significant improvements post-intervention, there was a clear trend towards a more balanced usage of both limbs, particularly evident in Axis 3, associated with vertical movement (p = 0.017). Additionally, a discernible correlation was observed between changes in the magnitude ratio derived from actigraphy data during CIMT (Δweek3-week1) and variations in traditional assessments pre- and post-intervention (ΔT0-T1), specifically the Assisting Hand Assessment grasp and release. Using tri-axial accelerometers has helped clarify the potential impacts of CIMT on children with UCP. The preliminary results suggest a possible link between actigraphy metrics taken during CIMT and the subsequent therapeutic outcomes determined by standardized tests.


[Leiomyomas of small bowel].
The analysis of our own country and foreign literature, devoted to the problems of clinical aspects, diagnosis and treatment of leiomyomas of the small bowel for the last 48 years has been carried out. The authors present their personal experience in follow-up of 6 cases of the small bowel leiomyomas, the main clinical manifestations of which being recurrent intestinal bleedings and in lesser degree--small bowel obstruction. The results of preoperative examination evidence that besides thorough physical, endoscopical and ultrasound examination the necessity may arise to use for diagnosis angiographic and computed-tomographic methods. The subject of special attention of each clinical physician should be the danger of malignant transformation of benign leiomyomatous neoplasms of the small bowel, which the authors encountered in 2 patients.


Corrected relative net protein ratio (CRNPR) method based on differences in rat and human requirements for sulfur amino acids.
The requirement for sulfur amino acids was calculated for growing rats fed 8% protein diets. The rat and human requirements for sulfur amino acids were compared and a correction factor was developed to reflect the differences. This correction factor was used to determine corrected relative net protein ratio (CRNPR) values for a number of potential meat extenders and their mixtures with beef. The methionine + cystine requirement of growing rats was estimated to be 4% of protein for 8% protein diets. The methionine + cystine requirement of rats was about 50% higher than that of humans (2.65% of protein). Based on this comparison (rat/human), a correction factor of 1.5 was developed to correct RNPR values of those protein products that were deficient in sulfur amino acids for rat growth. The CRNPR values of beef, casein, soybean protein products, pea concentrate, and peanut meal were 100, 100, 91-97, 75, and 73, respectively. Mixtures (50:50 protein basis) of beef with casein, soybean concentrate, soybean isolate, pea concentrate, peanut meal, rapeseed concentrate, rapeseed isolate, sunflower isolate, or wheat gluten were equal to beef in CRNPR values. The CRNPR method is a good predictor of protein quality for humans of those protein products that are deficient in sulfur amino acids.


Conductance in a bis-terpyridine based single molecular breadboard circuit.
Controlling charge flow in single molecule circuits with multiple electrical contacts and conductance pathways is a much sought after goal in molecular electronics. In this joint experimental and theoretical study, we advance the possibility of creating single molecule breadboard circuits through an analysis of the conductance of a bis-terpyridine based molecule (TP1). The TP1 molecule can adopt multiple conformations through relative rotations of 7 aromatic rings and can attach to electrodes in 61 possible single and multi-terminal configurations through 6 pyridyl groups. Despite this complexity, we show that it is possible to extract well defined conductance features for the TP1 breadboard and assign them rigorously to the underlying constituent circuits. Mechanically controllable break-junction (MCBJ) experiments on the TP1 molecular breadboard show an unprecedented 4 conductance states spanning a range 10 -2G0 to 10 -7G0. Quantitative theoretical examination of the conductance of TP1 reveals that combinations of 5 types of single terminal 2-5 ring subcircuits are accessed as a function of electrode separation to produce the distinct conductance steps observed in the MCBJ experiments. We estimate the absolute conductance for each single terminal subcircuit and its percentage contribution to the 4 experimentally observed conductance states. We also provide a detailed analysis of the role of quantum interference and thermal fluctuations in modulating conductance within the subcircuits of the TP1 molecular breadboard. Finally, we discuss the possible development of molecular circuit theory and experimental advances necessary for mapping conductance through complex single molecular breadboard circuits in terms of their constituent subcircuits.


Functional analysis and in vitro correction of splicing FAH mutations causing tyrosinemia type I.
Hereditary tyrosinemia type I (HT1) is a rare disease caused by a deficiency of fumarylacetoacetate hydrolase (FAH) in the tyrosine catabolic pathway, resulting mainly in hepatic alterations due to accumulation of the toxic metabolites fumarylacetoacetate, maleylacetoacetate and succinylacetone. We have characterized using minigenes four splicing mutations affecting exonic or intronic nucleotides of the FAH gene identified in two HT1 patients. Two of the mutations are novel, c.82-1G>A and c.913G>C and the other two have been previously associated with a splicing defect (c.836A>G and c.1062+5G>A). All mutations were confirmed to affect splicing in minigenes, resulting in exon skipping or activation of a cryptic splice site. We have analyzed the effect of different compounds known to modulate splicing (valproic acid, phenyl butyrate, M344, EIPA, and resveratrol) and the overexpression of splice factors of the SR protein family on the transcriptional profile of the mutant minigenes. For the c.836A>G mutation, a partial recovery of the correctly spliced transcript was observed. These results confirm the relevance of performing functional studies for mutations potentially affecting the splicing process and open the possibility of supplementary therapeutic approaches to diseases caused by splicing defects.


Cocaine-induced ANCA-associated renal disease: a case-based review.
Idiopathic antineutrophil cytoplasmic antibody (ANCA)-associated vasculitides (AAV) are a group of diseases that are often difficult to diagnose due to the wide range of clinical manifestations. Notably, renal involvement is a serious organ complication, which usually requires intensive immunosuppressive therapy and is prone to recurrence. In recent years, there has been some progress regarding the understanding of the pathogenesis of the diseases. It has been shown that both cocaine and levamisole, which is a common adulterant of cocaine, can trigger the formation of ANCAs and lead to the corresponding symptoms. We report two cases of AAV with different renal manifestations associated with cocaine consumption. Furthermore, we performed a review of the literature to identify, characterize and describe histologically documented cases of renal involvement in AAV, related to cocaine abuse. Cocaine/levamisole-induced vasculitis may, therefore, mimic idiopathic AAV. Although the detection of ANCA and anti-PR3 (proteinase 3, PR3) as well as anti-MPO antibodies (myeloperoxidase, MPO) are the serological hallmark of idiopathic AAV, certain clinical- and antibody constellations should lead to consideration of illicit drugs as inductors of the disease. Especially in young patients, certain serologic constellations (e.g., PR3 and MPO double positivity, positive antinuclear antibodies, low complement level, and positive testing for antiphospholipid antibodies), skin involvement, musculoskeletal symptoms and hematologic (anemia, leukopenia) affections should prompt testing for cocaine and levamisole consumption via urine drug testing. Treatment includes both immunosuppressive approaches and drug cessation but is difficult since many patients continue cocaine consumption.


Reaping the benefits of electronic medical record systems.
An electronic medical record system can provide benefits beyond the obvious functions of efficient and less labor-intensive scanning, archiving, retrieving, and printing of patient care information. The less tangible benefit of providing record access to several users simultaneously is difficult to quantify, but can enhance operations and improve the quality of patient care throughout a healthcare facility.


Administration of nerve growth factor-β to heifers with a pre-ovulatory follicle enhanced luteal formation and function and promoted LH release.
The objective of this study was to determine the effects of bovine nerve growth factor-β (NGF) on pre-ovulatory follicle vascular area, LH release, ovulation, and luteal function when administered systemically to heifers. Post-pubertal Holstein heifers (n = 12) received an intravaginal progesterone-releasing device (CIDR) and GnRH agonist (100 μg IM). The CIDR was removed 5 d later, and heifers were given dinoprost (25 mg IM) at CIDR removal and 24 h later, followed by a second dose of GnRH agonist 48 h later. Heifers were randomly assigned to treatments using a cross-over design. For example, heifers assigned to NGF (250 μg reconstituted in 12 mL PBS IM) in replicate 1 were assigned to control (12 mL PBS IM) in replicate 2. Transrectal ultrasonography was performed before treatment and repeated every 4 h up to 32 h to determine the pre-ovulatory follicle diameter, vascular area, and ovulation. Serum samples were obtained to assess LH concentrations during the periovulatory period and every 2 d post-ovulation for measuring progesterone concentrations. A subset of heifers had luteal biopsies performed on days 9 (n = 6 per treatment) and 14 (n = 6 per treatment) post-ovulation to count luteal cell numbers and measure relative mRNA abundance for steroidogenic and angiogenic enzymes and LH receptor. Treatment with NGF increased pre-ovulatory follicle diameter (P = 0.02) and serum LH concentrations (P = 0.03) but did not affect time to ovulation (P = 0.42). Heifers treated with NGF had increased serum progesterone concentrations in the subsequent luteal phase (P = 0.03), but no change in vascular area of the follicle (P = 0.16) or CL (P = 0.20). Heifers treated with NGF had a greater number of small luteal cells (P < 0.01) and a tendency for increased LH receptor (LHR) mRNA abundance in the CL (P = 0.10). There was also increased steroidogenic acute regulatory protein (STAR; P = 0.05) and a tendency for increased cytochrome P450 family 11 (CYP11A1; P = 0.10) mRNA abundance in the CL of NGF-treated heifers. There was decreased prostaglandin E2 synthase (PGES; P = 0.03) and its receptor (PGER; P = 0.05) mRNA abundance and a tendency for decreased cytochrome P450 family 17 subfamily A member 1 (CYP17A1; P = 0.08) and hydroxysteroid 17-beta dehydrogenase (HSD17B; P = 0.06) mRNA abundance in the CL of NGF-treated heifers. Administration of NGF improved CL function in heifers potentially as a result of increased LH release.


Scalp Cooling: Implementation of a Program at a Multisite Organization.
Chemotherapy-induced alopecia is a well-known side effect of some types of cancer treatments. With U.S. Food and Drug Administration clearance of two scalp cooling machines, patients with cancer now have the opportunity to minimize this hair loss. However, multiple barriers can exist for organizations when establishing a scalp cooling program. This article describes the experience of a large multisite organization that implemented a machine-based scalp cooling program. Nursing staff led an interprofessional team that addressed components of the program. As a result, eight sites within the authors' organization simultaneously began offering scalp cooling via machine using a single unified process. This approach was then successfully replicated one year later to prepare six additional sites to launch scalp cooling for other solid tumor types beyond breast cancer. Using a structured, collaborative, and interprofessional approach to the implementation of a scalp cooling program at the authors' institution allowed for standardization of care across sites. This approach can be replicated at other healthcare institutions.


Understanding droplet bridging in ionic liquid-based Pickering emulsions.
We have studied the unique bridging behavior of solid-stabilized oil-in-ionic liquid (IL) and water-in-ionic liquid emulsions with respect to particle concentration, particle size, and droplet phase using a confocal laser scanning microscope. The emulsions exhibited three morphology regimes: (1) single, sparingly covered droplets, (2) bridged clusters of droplets, and (3) fully covered droplets. The degree of bridging was directly proportional to the total potential bridging area which can be determined from the particle size and concentration. This type of emulsion diverges from much of the conventional wisdom of oil-water Pickering emulsions regarding the particle self-assembly onto droplet interfaces and liquid film stability. While the focus here is the bridging regime, we also report interesting observations, specifically, the deformed oil droplets and the transport of excess solid particles into the water droplets, in the fully covered droplet regime. The work identified new self-assembled particle structure and morphology in solid-stabilized emulsions.


Molecular conjugate-mediated gene transfer into isolated human kidneys.
Advances in systemic immunosuppressive therapy for solid organ transplantation have done little to decrease the percentage of allografts that eventually will develop chronic rejection. However, one of the promises of modern molecular biology includes the ability to introduce new genetic information into mammalian hosts. The ability to deliver genes and control their expression in the adult kidney has been described in appropriate animal models. Consequently, gene transfer technology represents a realistic therapeutic approach to modify the allogeneic kidney before engraftment in an effort to decrease the incidence of posttransplant dysfunction. To bridge the gap between animal studies and the clinical application of this technology, we report the first genetic transfection of isolated human kidneys under conditions of organ preservation. Polymerase chain reaction, reversed transcription polymerase chain reaction, and in situ hybridization techniques demonstrated that an adenovirus-polylysine-deoxyribonucleic acid (DNA) complex can be used to insert a complementary DNA expression vector encoding beta-galactosidase into the intact human kidney. Immunohistochemical and in situ enzymatic analyses determined further that gene delivery and expression were localized in proximal tubular epithelial cells. Consequently, targeting of genes to perturb mediators of the local inflammatory response may represent a rational therapeutic interventional strategy in chronic rejection of the kidney.


Hyperactivation of stallion sperm is required for successful in vitro fertilization of equine oocytes.
Capacitation is a complex and not well-understood process that encompasses all the molecular changes sperm must undergo to successfully fertilize an oocyte. In vitro fertilization has remained elusive in the horse, as evidenced by low in vitro fertilization (IVF) rates (0%-33%); moreover, only two foals have ever been produced using IVF. Incubation of stallion sperm in modified Whittens supplemented with bovine serum albumin and sodium bicarbonate yielded significant rates of time-dependent protein tyrosine phosphorylation and induced acrosomal exocytosis, consistent with capacitation. The objective of this study was to characterize stallion sperm hyperactivation and to test whether hyperactivation of capacitated sperm supported equine IVF. Treatment of sperm with procaine, an anesthetic shown to induce hyperactivation in other mammalian species, resulted in the decrease of three motility variables indicative of hyperactivation: straight line velocity (P = 0.029), straightness (P = 0.001), and linearity (P = 0.002). We demonstrated that procaine-induced hyperactivation was not regulated by changes in protein tyrosine phosphorylation and that it did not induce acrosomal exocytosis in capacitated sperm compared with calcium ionophore (P > 0.05), similar to findings in the bovine. Most notably, by coupling our capacitating conditions with the induction of hyperactivation using procaine, we have achieved the novel result of substantial and reproducible percentages of fertilized mare oocytes (60.7%) in our IVF experiments. Conversely, sperm incubated in capacitating conditions but not treated with procaine did not fertilize (0%). These results support the hypothesis that capacitation and hyperactivation are required for successful IVF in the equine.


Genetic analysis of cytoprotective functions supported by graded expression of Keap1.
Keap1 regulates Nrf2 activity in response to xenobiotic and oxidative stresses. Nrf2 is an essential regulator of cytoprotective genes. Keap1-null mice are lethal by weaning age due to malnutrition caused by severe hyperkeratosis of the upper digestive tract. Analysis of Keap1::Nrf2 double mutant mice revealed that currently recognizable phenotypes of Keap1-null mice are all attributable to constitutive activation of Nrf2. We previously reported that hepatocyte-specific Keap1 knockout (Keap1(flox/-)::Albumin-Cre) mice are viable and more resistant to acute toxicity of acetaminophen (APAP). In the current study, we found that the floxed Keap1 allele is hypomorphic and that Keap1 expression was decreased in all examined tissues of Keap1(flox/-) mice. Taking advantage of the hypomorphic phenotype of Keap1(flox/-) mice, we examined the effects of graded reduction of Keap1 expression in adult mice. When challenged with APAP, Keap1(flox/-) mice were more protected from mortality than wild-type and even Keap1(flox/-)::Albumin-Cre mice. In contrast, a decrease in Keap1 levels to less than 50% resulted in increased mortality in a study of 2-year-old mice. These results support our contention that the benefits of Nrf2 activation in acute toxicity are hormetic and that constitutive Nrf2 activation beyond a certain threshold is rather disadvantageous to long-term survival.


Medial plantar nerve conduction studies in healthy controls and diabetics.
To collect a reference material of the medial plantar nerve action potential, to test intra/interobserver reliability in healthy controls and to apply the test to a group of patients with diabetes mellitus. 98 healthy controls and 50 patients with diabetes mellitus were included. The medial plantar nerve was stimulated orthodromically and recorded with a surface electrode. In the patient group, NCS of motor and sensory nerves and quantitative sensory testing were also performed. Responses of the medial plantar nerve were obtained from all controls except from one aged 72. Amplitude decreased with age (r=-0.68, p<0.0001). Intra/interobserver reliability was acceptable. 52% of the patients had abnormal overall NCS classification. Forty-eight percent had delayed tibial F-response latency. The medial plantar NCS were abnormal in 59% of the cases (47% abnormal NAP amplitude and 39% reduced CV), 59% of those with abnormal NCS had symptoms of sensory polyneuropathy. Only 24% had abnormal sural amplitude. Cold perception threshold was abnormal in more patients (30%) than warmth perception threshold (14%). Responses were easily obtained in controls under 70 years. In diabetics the amplitudes of the medial plantar nerve were abnormal more often than in the sural nerve. The medial plantar nerve response is reliable in patients under 70 years, and intra/interobserver reliability is acceptable.


Dendritic potassium channel dysfunction may contribute to dendrite degeneration in spinocerebellar ataxia type 1.
Purkinje neuron dendritic degeneration precedes cell loss in cerebellar ataxia, but the basis for dendritic vulnerability in ataxia remains poorly understood. Recent work has suggested that potassium (K+) channel dysfunction and consequent spiking abnormalities contribute to Purkinje neuron degeneration, but little attention has been paid to how K+ channel dysfunction impacts dendritic excitability and the role this may play in the degenerative process. We examined the relationship between K+ channel dysfunction, dendritic excitability and dendritic degeneration in spinocerebellar ataxia type 1 (SCA1). Examination of published RNA sequencing data from SCA1 mice revealed reduced expression of several K+ channels that are important regulators of excitability in Purkinje neuron dendrites. Patch clamp recordings in Purkinje neurons from SCA1 mice identified increased dendritic excitability in the form of enhanced back-propagation of action potentials and an increased propensity to produce dendritic calcium spikes. Dendritic excitability could be rescued by restoring expression of large-conductance calcium-activated potassium (BK) channels and activating other K+ channels with baclofen. Importantly, this treatment combination improves motor performance and mitigates dendritic degeneration in SCA1 mice. These results suggest that reduced expression of K+ channels results in persistently increased dendritic excitability at all stages of disease in SCA1, which in turn may contribute to the dendritic degeneration that precedes cell loss.


SNPs across time and space: population genomic signatures of founder events and epizootics in the House Finch (Haemorhous mexicanus).
Identifying genomic signatures of natural selection can be challenging against a background of demographic changes such as bottlenecks and population expansions. Here, we disentangle the effects of demography from selection in the House Finch (Haemorhous mexicanus) using samples collected before and after a pathogen-induced selection event. Using ddRADseq, we genotyped over 18,000 SNPs across the genome in native pre-epizootic western US birds, introduced birds from Hawaii and the eastern United States, post-epizootic eastern birds, and western birds sampled across a similar time span. We found 14% and 7% reductions in nucleotide diversity, respectively, in Hawaiian and pre-epizootic eastern birds relative to pre-epizootic western birds, as well as elevated levels of linkage disequilibrium and other signatures of founder events. Despite finding numerous significant frequency shifts (outlier loci) between pre-epizootic native and introduced populations, we found no signal of reduced genetic diversity, elevated linkage disequilibrium, or outlier loci as a result of the epizootic. Simulations demonstrate that the proportion of outliers associated with founder events could be explained by genetic drift. This rare view of genetic evolution across time in an invasive species provides direct evidence that demographic shifts like founder events have genetic consequences more widespread across the genome than natural selection.


Selective Expression of the MAPK Phosphatase Dusp9/MKP-4 in Mouse Plasmacytoid Dendritic Cells and Regulation of IFN-β Production.
Plasmacytoid dendritic cells (pDCs) efficiently produce large amounts of type I IFN in response to TLR7 and TLR9 ligands, whereas conventional DCs (cDCs) predominantly secrete high levels of the cytokines IL-10 and IL-12. The molecular basis underlying this distinct phenotype is not well understood. In this study, we identified the MAPK phosphatase Dusp9/MKP-4 by transcriptome analysis as selectively expressed in pDCs, but not cDCs. We confirmed the constitutive expression of Dusp9 at the protein level in pDCs generated in vitro by culture with Flt3 ligand and ex vivo in sorted splenic pDCs. Dusp9 expression was low in B220(-) bone marrow precursors and was upregulated during pDC differentiation, concomitant with established pDC markers. Higher expression of Dusp9 in pDCs correlated with impaired phosphorylation of the MAPK ERK1/2 upon TLR9 stimulation. Notably, Dusp9 was not expressed at detectable levels in human pDCs, although these displayed similarly impaired activation of ERK1/2 MAPK compared with cDCs. Enforced retroviral expression of Dusp9 in mouse GM-CSF-induced cDCs increased the expression of TLR9-induced IL-12p40 and IFN-β, but not of IL-10. Conditional deletion of Dusp9 in pDCs was effectively achieved in Dusp9(flox/flox); CD11c-Cre mice at the mRNA and protein levels. However, the lack of Dusp9 in pDC did not restore ERK1/2 activation after TLR9 stimulation and only weakly affected IFN-β and IL-12p40 production. Taken together, our results suggest that expression of Dusp9 is sufficient to impair ERK1/2 activation and enhance IFN-β expression. However, despite selective expression in pDCs, Dusp9 is not essential for high-level IFN-β production by these cells.


Spinal process landmark as a predicting factor for difficult epidural block: a prospective study in Taiwanese patients.
Although epidural anesthesia is a common practice in neuraxial blockade, difficult access to the epidural space is a frequent problem in operating theaters. We designed this study of epidural blocks to determine if the spinal landmark grading system is valuable in predicting a difficult epidural block. Before the epidural block, we collected the following data: demographics, body habitus (normal, thin, obese, pregnant), spinal anatomy (normal, deformed), spinal level (lumbar, thoracic), and spinal landmark grade (grade 1: spinous processes visible; grade 2: spinous processes not seen but easily palpated; grade 3: spinous processes not seen and not palpated but the interval between them is palpated as a low landmark under the thumb; grade 4: other). We performed all 848 epidural blocks initially using a midline approach and an 18-gauge Touhy needle. We evaluated the technical difficulty of the epidural block using three methods: whether the epidural block was accomplished at the spinal level (first-level success); the total number of attempts at skin puncture (attempts-S); and total number of attempts to change ligament puncture direction (attempts-L) required to complete the epidural block. Of all examined factors, spinal landmark grade correlated best with technical difficulty as measured by all three methods. Deformed spinal anatomy and body habitus both correlated with difficulty, merely from the total numbers of attempts (attempts-S and attempts-L). Thoracic epidurals were more difficult than lumbar epidurals by all three measures of difficulty. We concluded that this spinal landmark grading system is valuable in predicting a difficult epidural block and advocate its use as a predictor by anesthesiologists.


Seizures in Fragile X Syndrome: Associations and Longitudinal Analysis of a Large Clinic-Based Cohort.
Fragile X syndrome (FXS), the most common inherited cause of intellectual disability, learning disability, and autism spectrum disorder, is associated with an increased prevalence of certain medical conditions including seizures. The goal of this study was to better understand seizures in individuals with FXS using the Fragile X Online Registry with Accessible Research Database, a multisite observational study initiated in 2012 involving FXS clinics in the Fragile X Clinic and Research Consortium. Seizure data were available for 1,607 participants, mostly male (77%) and white (74.5%). The overall prevalence of at least one seizure was 12%, with this rate being significantly higher in males than females (13.7 vs. 6.2%, p < 0.001). As compared to individuals with FXS without seizures, those with seizures were more likely to have autism spectrum disorder, current sleep apnea, later acquisition of expressive language, more severe intellectual disability, hyperactivity, irritability, and stereotyped movements. The mean age of seizure onset was 6.4 (SD 6.1) years of age with the great majority (>80%) having onset of seizures which was before 10. For those with epilepsy, about half (52%) had seizures for more than 3 years. This group was found to have greater cognitive and language impairment, but not behavioral disruptions, compared with those with seizures for <3 years. Antiepileptic drugs were more often used in males (60.6%) than females (34.8%), and females more often required more than one medication. The most commonly used anticonvulsants were oxcarbazepine, valproic acid, lamotrigine, and levetiracetam. The current study is the largest and first longitudinal study ever conducted to describe seizures in FXS. Overall, this study confirms previous reports of seizures in FXS and extends previous findings by further defining the cognitive and behavioral phenotype of those with epilepsy in FXS. Future studies should further investigate the natural history of seizures in FXS and the characteristics of seizures in FXS in adulthood.


[Contribution of the Rorschach test to the differential diagnosis of impotence].
Attention is called to the clinical problem of masked depressions and depressive equivalents. In certain situations, the "impotence" symptom can be classified sonographically among cyclothymic disturbances. The Rorschach test has been used to study some typical cases and it is stressed that a psychodiagnostic investigation may be useful in the differential diagnosis of the clinical syndrome.


A generic framework for spatial quantitative risk assessments of infectious diseases: Lumpy skin disease case study.
The increase in availability of spatial data and the technological advances to handle such data allow for subsequent improvements in our ability to assess risk in a spatial setting. We provide a generic framework for quantitative risk assessments of disease introduction that capitalizes on these new data. It can be adopted across multiple spatial scales, for any pathogen, method of transmission or location. The framework incorporates the risk of initial infection in a previously uninfected location due to registered movement (e.g., trade) and unregistered movement (e.g., daily movements of wild animals). We discuss the steps of the framework and the data required to compute it. We then outline how this framework is applied for a single pathway using lumpy skin disease as a case study, a disease which had an outbreak in the Balkans in 2016. We calculate the risk of initial infection for the rest of Europe in 2016 due to trade. We perform the risk assessment on 3 spatial scales-countries, regions within countries and individual farms. We find that Croatia (assuming no vaccination occurred) has the highest mean probability of infection, with Italy, Hungary and Spain following. Including import detection of infected trade does reduce risk but this reduction is proportionally lower for countries with highest risk. The risk assessment results are consistent across the spatial scales, while in addition, at the finer spatial scales, it highlights specific areas or individual locations of countries on which to focus surveillance.


[THE DEVELOPMENT OF TECHNIQUE OF DIAGNOSTIC OF VUVLVOVAGINAL CANDIDIASIS BASED ON QUANTITATIVE MULTIPLEX POLYMERASE CHAIN REACTION].
The technique is developed including corresponding kit of reagents "AmpliSens Florocenose/Candida-FL", to be applied in laboratory diagnostic of vuvlvovaginal candidiasis. The technique is based on polymerase chain reaction in real-time and permits identifying and quantitatively estimate in biological material content of five most prevalent agents of candidiasis: Candida albicans, C. glabrata, C. krusei, C. paprapsilosis, C. tropicalis. The experimental determination of analytical characteristics of developed technique was carried out. The analytical sensitivity amounted to 100 GE/mL (genome equivalent in 1 mL), linear range -from 200 to 2 x 10 7 GE/mL for every specie of detected Candida. The comparison of quantitative results of detection of content of Candida spp using the developed technique with results of inoculation demonstrated their mutual matching. In logarithm presentation the relation coefficient of lg (GE/mL) to lg (CFU/mL) amounted to 1.2. The correlation of results characterized by coefficient R 2 equal 0.76.


Reference values for Lactate Pro 2™ in fetal blood sampling during labor: a cross-sectional study.
Lactate Pro™ (LP1) is the only lactate meter evaluated for fetal scalp blood sampling (FBS) in intrapartum use. The reference values for this meter are: normal value <4.2 mmol/L, preacidemia 4.2-4.8 mmol/L, and acidemia >4.8 mmol/L. The production of this meter has been discontinued. An updated version, Lactate Pro 2TM (LP2), has been launched and is shown to be differently calibrated. The aims of the study were to retrieve a conversion equation to convert lactate values in FBS measured with LP2 to an estimated value if using LP1 and to define reference values for clinical management when using LP2. A cross-sectional study was conducted at a university hospital in Sweden. A total of 113 laboring women with fetal heart rate abnormalities on cardiotocography (CTG) had FBS carried out. Lactate concentration was measured bedside with both LP1 and LP2 from the same blood sample capillary. A linear regression model was constructed to retrieve a conversion equation to convert LP2 values to LP1 values. LP2 measured higher values than LP1 in all analyses. We found that 4.2 mmol/L with LP1 corresponded to 6.4 mmol/L with LP2. Likewise, 4.8 mmol/L with LP1 corresponded to 7.3 mmol/L with LP2. The correlation between the analyses was excellent (Spearman's rank correlation, r=0.97). We recommend the following guidelines when interpreting lactate concentration in FBS with LP2: <6.4 mmol/L to be interpreted as normal, 6.4-7.3 mmol/L as preacidemia indicating a follow-up FBS within 20-30 min, and >7.3 mmol/L as acidemia indicating intervention.


Efficient all-optical 2R regeneration using self-phase modulation in bidirectional fiber configuration.
An efficient wavelength-conversion-free all-optical 2R regenerator is proposed. The regeneration is based on power-dependent spectrum broadening in normal-dispersion fiber and subsequent off-centered filtering. Twofold regeneration is performed in a bidirectional configuration where the signal is transmitted twice along the fiber in opposite directions. Experiment at 10Gb/s shows no penalty arising from the bidirectional high-power signal transmission in the fiber and demonstrates strong improvement of extinction ratio of the input signal by the regenerator.


Proteaphagy in Mammalian Cells Can Function Independent of ATG5/ATG7.
The degradation of intra- and extracellular proteins is essential in all cell types and mediated by two systems, the ubiquitin-proteasome system (UPS) and the autophagy-lysosome pathway. This study investigates the changes in autophagosomal and lysosomal proteomes upon inhibition of proteasomes by bortezomib (BTZ) or MG132. We find an increased abundance of more than 50 proteins in lysosomes of cells in which the proteasome is inhibited. Among those are dihydrofolate reductase (DHFR), β-Catenin and 3-hydroxy-3-methylglutaryl-coenzym-A (HMGCoA)-reductase. Because these proteins are known to be degraded by the proteasome they seem to be compensatorily delivered to the autophagosomal pathway when the proteasome is inactivated. Surprisingly, most of the proteins which show increased amounts in the lysosomes of BTZ or MG132 treated cells are proteasomal subunits. Thus an inactivated, non-functional proteasome is delivered to the autophagic pathway. Native gel electrophoresis shows that the proteasome reaches the lysosome intact and not disassembled. Adaptor proteins, which target proteasomes to autophagy, have been described in Arabidopsis, Saccharomyces and upon starvation in mammalians. However, in cell lines deficient of these proteins or their mammalian orthologues, respectively, the transfer of proteasomes to the lysosome is not impaired. Obviously, these proteins do not play a role as autophagy adaptor proteins in mammalian cells. We can also show that chaperone-mediated autophagy (CMA) does not participate in the proteasome delivery to the lysosomes. In autophagy-related (ATG)-5 and ATG7 deficient cells the delivery of inactivated proteasomes to the autophagic pathway was only partially blocked, indicating the existence of at least two different pathways by which inactivated proteasomes can be delivered to the lysosome in mammalian cells.


Effects of Alpha-Lipoic Acid Supplementation on Cardiovascular Disease Risk Factors in β-Thalassemia Major Patients: A Clinical Trial Crossover Study.
Thalassemia is one of the most common genetic diseases, and cardiovascular disease (CVD) has been considered as the leading cause of mortality in more than 50% of β-thalassemia patients. The aim of this study was to determine the effects of alpha-lipoic acid (ALA) on CVD risk factors in β-thalassemia major patients. Twenty β-thalassemia major patients participated in this randomized crossover clinical trial study. Participants were randomly assigned to ALA (600 mg/day) or placebo groups for two 8-wk interventions that were separated by a 3-wk washout period. The CVD risk factors including serum osteoprotegerin (OPG), homocysteine, lipoprotein-associated phospholipase A2 and trimethylamine N-oxide were measured at the beginning and the end of each intervention phase according to the standard protocol. Serum OPG reduced significantly in the ALA group in all participants (5.38 ± 2.79 to 3.27 ± 2.43 ng/mL, P= .003) and in the male subgroup (5.24 ± 2.56 to 3.13 ± 2.5 ng/mL, P= .015); this reduction was significant in comparison with the placebo group (P= .013). The changes in other CVD risk factors were not significant. The results of this study showed that after 8-wk of ALA consumption, the serum OPG reduced significantly in β-thalassemia major patients. Therefore, controlling the serum OPG level with ALA consumption can be an important complementary therapeutic option to prevent the progression of CVD in β-thalassemia major patients.


[Evaluation of the antimicrobial susceptibility profile of microorganisms isolated from urine in the region of Vale do Sousa and Tâmega].
The urinary tract is one of the more common sites of bacterial infections, especially in women. Urinary infection can be defined as an infection of urinary tract structures which occurs, generally, as a consequence of the presence or colonization by urine bacteria. The aim of this study was to determine the etiology of urinary tract infections and their susceptibility to antimicrobial agents in the region of Vale do Sousa and Tâmega. From February 2008 to January 2009, 18653 urine cultures were analyzed. From these cultures 1037 were positive. From this total of positive cultures, 18,3% were from males and 81.7% were from females. In bacteriological positive tests, 23 different strains of microorganisms were found. It was verified that the most frequent microorganism was Escherichia coli, followed by Proteus mirabilis, Klebsiella pneumoniae, Enterococcus faecalis and Pseudomonas aeruginosa. For antimicrobial susceptibility it was verified that Escherichia coli showed low susceptibility to amoxicillin and cotrimoxazole. Proteus mirabilis showed good susceptibility to cefotaxime and low to cotrimoxazole, amoxicillin and ciprofloxacin. For Klebsiella pneumoniae was found only a reasonable susceptibility to gentamicin and Enterococcus faecalis showed amoxicillin susceptibility. Pseudomonas aeruginosa showed low susceptibility to all antibiotics analyzed with the exception of the combination piperacillin/tazobactam. In conclusion, this study shows that Escherichia coli, Proteus mirabilis and Klebsiella pneumoniae are the three main microorganisms that causes urinary infections in the region of Vale do Sousa and Tâmega. Therefore, the antimicrobial empirically used must have a spectrum against enterobacteria, because they are the most likely to be present in urinary tract infections acquired in the community. A periodic analysis of the susceptibility profile should be performed over time for each region in order to help in the beginning of the empirical antimicrobial treatment.


Loop F of the GABAA receptor alpha subunit governs GABA potency.
Gamma-amino butyric acid (GABA) is an abundant neurotransmitter in the CNS. GABAergic interneurons orchestrate pyramidal neurons in the cerebral cortex, and thus control learning and memory. Ionotropic receptors for GABA (GABAAR) are heteropentameric complexes of α, β and γ integral membrane-protein subunits forming Cl- -channels operated by GABA, which are vital for brain function and are important drug targets. However, knowledge on how GABAAR bind GABA is controversial. Structural biology versus functional modelling combined with site-directed mutagenesis suggest markedly different roles for loop F of the extracellular domain of the α-subunit when complexed with GABA. Here, we report that contrary to the results of structural studies, loop F of the α-subunit controls the potency of GABA on GABAAR. We examined the effect of replacing a short, variable segment of loop F of the GABAA α5-subunit with the corresponding segment of the α2-subunit (GABAA5_LF2) and vice versa (GABAA2-LF5). When compared with their respective wild-type counterparts, GABAA5_LF2 receptors displayed enhanced sensitivity towards GABA, whilst in GABAA2-LF5 sensitivity was diminished. Mice homozygous for the genetic knock-in of the GABAA5_LF2 subunit showed a marked deficit in long- but not short-term object recognition memory. Working memory in place learning, spontaneous alternation and the rewarded T-maze were all normal. The deficit in long-term recognition memory was reversed by an α5-GABAA negative allosteric modulator compound. The data show that loop F governs GABA potency in a receptor isoform-specific manner in vitro. Moreover, this mechanism of ligand recognition appears to be operative in vivo and impacts cognitive performance.


Does cAMP response element-binding protein have a pivotal role in hippocampal synaptic plasticity and hippocampus-dependent memory?
Previous studies addressing the role of the transcription factor cAMP response element-binding protein (CREB) in mammalian long-term synaptic plasticity and memory by gene targeting were compromised by incomplete deletion of the CREB isoforms. Therefore, we generated conditional knock-out strains with a marked reduction or complete deletion of all CREB isoforms in the hippocampus. In these strains, no deficits could be detected in lasting forms of hippocampal long-term potentiation (LTP) and long-term depression (LTD). When tested for hippocampus-dependent learning, mutants showed normal context-dependent fear conditioning. Water maze learning was impaired during the early stages, but many mutants showed satisfactory scores in probe trials thought to measure hippocampus-dependent spatial memory. However, conditioned taste aversion learning, a putatively hippocampus-independent memory test, was markedly impaired. Our data indicate that in the adult mouse brain, loss of CREB neither prevents learning nor substantially affects performance in some hippocampus-dependent tasks. Furthermore, it spares LTP and LTD in paradigms that are sensitive enough to detect deficits in other mutants. This implies either a species-specific or regionally restricted role of CREB in the brain and/or a compensatory upregulation of the cAMP response element modulator (CREM) and other as yet unidentified transcription factors.


An evaluation of the cost-effectiveness of omalizumab for the treatment of severe allergic asthma.
Omalizumab is the first licensed anti-immunoglobulin (Ig) E antibody shown to be effective for treatment of allergic (IgE-mediated) asthma. Recent international guidelines recommend omalizumab as add-on treatment to fixed dose inhaled corticosteroid (ICS) and long-acting beta(2)-agonist (LABA) combination therapy. However, omalizumab is more expensive than other current asthma treatments and health and reimbursement authorities are increasingly demanding evidence of economic benefit to support pricing and formulary listing. The aims of this article are to (i) summarize data on the human and economic burden of severe asthma, (ii) summarize the efficacy data obtained for omalizumab in clinical trials in patients with inadequately controlled severe persistent allergic asthma despite high-dose ICS plus a LABA, and (iii) discuss the cost-effectiveness evidence published for omalizumab in this patient population. A wealth of evidence exists highlighting that the health, economic and societal burden of asthma is considerable and is highly skewed towards patients with severe asthma, particularly when asthma is inadequately controlled. Omalizumab is clinically beneficial in patients with severe persistent allergic asthma despite high-dose ICS plus a LABA, particularly in a subgroup of patients who respond to therapy. In patients who respond to therapy, the cost-effectiveness of omalizumab compares well with other biologic treatments for chronic illness.


Iridocorneal adhesions in posterior polymorphous dystrophy.
The variability of clinical expression in posterior polymorphous dystrophy is illustrated with emphasis on the occurrence of iridocorneal adhesions. These are believed to be synechiae and not Rieger's anomaly. The occurrence of glass-membrane-like material extending onto the iris from the cornea, causing synechiae and pupillary ectropion, is documented.


Hirschsprung disease and more: dysregulation of ERBB2 and ERBB3.
The enteric nervous system mediates reflexes independently of the brain and spinal cord and transmits signals bidirectionally between the gut and the brain. Hirschsprung disease and chronic intestinal pseudo-obstruction (CIPO) and pediatric CIPO are examples of congenital defects that impair gastrointestinal motility. In this issue of the JCI, Thuy-Linh Le et al. analyzed eight patients with defects in tissue that arose from the neural crest. The patients carried homozygous or heterozygous variants in ERBB3 or ERBB2, which encode transmembrane epidermal growth factor receptors that bind neuroregulin 1 (NRG1). Notably, the genetic variants resulted in loss of function with decreased expression or aberrant phosphorylation of the ERBB3/ERBB2 receptors. Experiments using mice revealed that Erbb3 and Erbb2 were expressed in enteric neuronal progenitor cells. This study is an outstanding example of descriptive observation that begs for mechanistic exploration to reveal precisely how the NRG1/ERBB3/ERBB2 pathway influences ENS development.


Gamma Knife radiosurgery for intracranial meningiomas: Do we need to treat the dural tail? A single-center retrospective analysis and an overview of the literature.
The dural tail (DT) has been described as a common feature in meningiomas. There is a great variation of tumor invasion and extent of tumor cells in the DT. Therefore, the necessity to include the whole DT in Gamma Knife radiosurgery is not clear, since inclusion increases the target volume and therefore increases the risk of complications. In this analysis, we evaluated whether the complete tail should be included as part of the target in Gamma Knife radiosurgery for meningiomas. Between June 2002 and December 2010, Gamma Knife radiosurgery was performed in 160 patients with 203 meningiomas with a DT. In 105 tumors, the diagnosis was based on magnetic resonance imaging (MRI) characteristics, and in 98 tumors, the diagnosis was confirmed by histopathologic examination after surgery. The median volume of the tumors was 3.55 cc. All tumors were treated with Gamma Knife radiosurgery with a median prescribed dose of 13 Gy (range 11-15), resulting in a median marginal dose of 11 Gy (range 10-15). Only the part of the DT closely related to the tumor mass was included in the target. The median follow-up period was 41 months (range 12-123). In image-based meningiomas, the overall local control rate was 96.2% with 2- and 5-year control rates of 98.0% and 95.1%, respectively. In WHO grade I tumors, the overall local control rate was 85.9% with 2- and 5-year control rates of 94.5% and 88.0%, respectively. The overall local control rate in World Health Organization (WHO) grade II tumors was 70.6% with control rates of 83.4% and 64.4% after 2 and 5 years, respectively. The growth of all new tumors was found in the radiation target area. No tumor growth was observed in the part of the DT that had been excluded from the target volume. We found in this study that routinely excluding the DT from the target does not lead to out-of-field tumor progression. Given the possibility that the DT is infiltrated with tumor cells, regular follow-up is needed.


Evidence for Ig Light Chain Isotype Exclusion in Shark B Lymphocytes Suggests Ordered Mechanisms.
Unlike most vertebrates, the shark IgL gene organization precludes secondary rearrangements that delete self-reactive VJ rearranged genes. Nurse sharks express four L chain isotypes, κ, λ, σ, and σ-2, encoded by 35 functional minigenes or clusters. The sequence of gene activation/expression and receptor editing of these isotypes have not been studied. We therefore investigated the extent of isotypic exclusion in separated B cell subpopulations. Surface Ig (sIg)κ-expressing cells, isolated with mAb LK14 that recognizes Cκ, carry predominantly nonproductive rearrangements of other L chain isotypes. Conversely, after depletion with LK14, sIgM+ cells contained largely nonproductive κ and enrichment for in-frame VJ of the others. Because some isotypic inclusion was observed at the mRNA level, expression in the BCR was examined. Functional λ mRNA was obtained, as expected, from the LK14-depleted population, but was also in sIgκ+ splenocytes. Whereas λ somatic mutants from the depleted sample displayed evidence of positive selection, the λ genes in sIgκ+ cells accumulated bystander mutations indicating a failure to express their products at the cell surface in association with the BCR H chain. In conclusion, a shark B cell expresses one L chain isotype at the surface and other isotypes as nonproductive VJ, sterile transcripts, or in-frame VJ whose products may not associate with the H chain. Based on the mRNA content found in the B cell subpopulations, an order of L chain gene activation is suggested as: σ-2 followed by κ, then σ and λ.


Tellegacy: An Intergenerational Wellness and Health Promotion Project to Reduce Social Isolation and Loneliness in Older Adults: A Feasibility Study.
Emerging research demonstrates that social isolation and loneliness are linked to significant physical and mental health conditions. To address these concerns, the Tellegacy program was developed as an intergenerational health-promoting intervention to ameliorate older adult social isolation and loneliness in an effort to increase wellness. The purpose of this study was to reflect on testing of the Tellegacy program as a behavioral intervention. University students trained in goal setting, mindfulness, and listening strategies were paired with 11 older adults in the northern Midwest area via weekly in-person and phone conversations. Oral reminiscence therapies were used and books containing their stories were given to the older adults after participation. Older adults were surveyed using the University of California Los Angeles Loneliness Scale, Satisfaction of Life Scale, and patient health questionnaire-9 (PHQ-9) scale to elucidate the effectiveness of the intervention. Improved scores in loneliness, satisfaction of life, and PHQ-9 demonstrated favorable improvements in older adults. Additional benefits for the student Legacy Builder were revealed from self-reported changes. This suggests the potential benefits of structured encounters between trained students and isolated or lonely older adults. The Tellegacy intergenerational feasibility program warrants further studies to fully demonstrate its impact on health outcomes.


Thyroid Hormone Therapy and Risk of Thyrotoxicosis in Community-Resident Older Adults: Findings from the Baltimore Longitudinal Study of Aging.
Both endogenous and exogenous thyrotoxicosis has been associated with atrial fibrillation and low bone mineral density. Therefore, this study investigated the risk factors associated with prevalent and incident thyrotoxicosis and the initiation of thyroid hormone therapy in a healthy, aging cohort. A total of 1450 ambulatory community volunteer participants in the Baltimore Longitudinal Study of Aging examined at the NIA Clinical Research Unit in Baltimore, MD, have undergone longitudinal monitoring of serum thyrotropin (TSH) and thyroid hormone (free thyroxine and free triiodothryonine) levels as well as medication use every one to four years, depending on age, between 2003 and 2014. The prevalence of low TSH was 9.6% for participants on thyroid hormone and 0.8% for nontreated individuals (p < 0.001). New cases occurred at a rate of 17.7/1000 person-years of exposure to thyroid hormone therapy [CI 9-32/1000] and 1.5/1000 person-years in the unexposed population [CI 0.7-2.9/1000]. Women were more likely to be treated and more often overtreated than men were. The adjusted hazard ratio (HR) for thyrotoxicosis between treated and untreated women was 27.5 ([CI 7.2-105.4]; p < 0.001) and 3.8 for men ([CI 1.2-6.3]; p < 0.01). White race/ethnicity and older age were risk factors for thyroid hormone therapy but not overtreatment. Body mass index was not associated with starting therapy (HR = 1.0). Thyroid hormone initiation was highest among women older than 80 years of age (3/100 person-years). For one-third of treated participants with follow-up data, overtreatment persisted at least two years. Iatrogenic thyrotoxicosis accounts for approximately half of both prevalent and incident low TSH events in this community-based cohort, with the highest rates among older women, who are vulnerable to atrial fibrillation and osteoporosis. Physicians should be particularly cautious in treating subclinical hypothyroidism in elderly women in light of recent studies demonstrating no increased risk of cardiovascular morbidity or death for individuals with elevated TSH levels <10 mIU/L.