Induction of the enzyme aldose reductase in a lens epithelial cell line from a transgenic mouse.
A lens epithelial cell line established from a transgenic mouse synthesizes high levels of the enzyme aldose reductase which converts sugars to polyols. This enzyme has been implicated in the formation of sugar cataracts in animals and with diabetic complications in man. The mouse aldose reductase has been characterized and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis has an apparent molecular mass of 38,000, similar to the enzyme in rat and man. The cellular enzyme is inhibited by two aldose reductase inhibitors: Sorbinil (IC50 = 1.8 X 10(-7) M) and Alcon 1576 (IC50 = 7.8 X 10(-8) M). The amount and the specific activity of the aldose reductase can be further increased in the cells by raising the osmolarity of the medium to 500 mOSM. Although the amount of aldose reductase is increased approximately sevenfold under these conditions, alpha-crystallin, one of the main lens specific proteins, remained at about the same concentration. No detectable increase in sorbitol was found within the cells, in contrast to published reports on renal cells in which this polyol increases under similar hyperosmotic conditions; however, in the lens cells there was a five-fold increase in the inositol content, suggesting that this polyol rather than sorbitol may be used to compensate for some of the changes in the osmolarity. The induction of the enzyme aldose reductase without the apparent accumulation of its product suggests a complex mechanism for osmoregulation in the lens cells.


Anorexia nervosa in a patient with systemic lupus erythematosus.
Anorexia nervosa has not been described in patients with systemic lupus erythematosus (SLE). The case of a patient with SLE who developed the classical features of anorexia nervosa is described. Although an impressive interplay between psychosocial and cultural factors was present, the possibility that this syndrome was produced by active CNS vasculitis remained.


Role of blood urea nitrogen in predicting the post-discharge prognosis in elderly patients with acute decompensated heart failure.
Blood urea nitrogen (BUN) is a surrogate marker for neurohormonal activation, but the association between BUN and the post-discharge prognosis in elderly patients with acute decompensated heart failure (ADHF) is not well defined. We explored the association between BUN and post-discharge all-cause mortality in 652 elderly patients (73.9 ± 7.8 yr) with ADHF. All patients were followed for a mean duration of 32 months (12-69 months). BUN was analyzed both as a continuous variable and according to two categories: low BUN group (BUN < 15.35 mmol/L, N = 361) and high BUN group (BUN ≥ 15.35 mmol/L, N = 291). The risk of all-cause mortality increased by 1.6% per 1 mmol/L increase in BUN concentration when BUN was used as a continuous variable [hazard ratio (HR): 1.016, 95% confidence interval (CI): 1.006-1.026, p = 0.002]. BUN maintained an independent and significant positive correlation with all-cause mortality as a categorical variable (HR: 1.355, 95% CI: 1.023-1.794, p = 0.034 for the high BUN group). The BUN C-statistic for predicting all-cause mortality was 0.624 (95% CI: 0.585-0.661). The cut-off value for BUN was 15.35 mmol/L with sensitivity of 0.58 and specificity of 0.63. The prognostic performance of BUN was similar to brain natriuretic peptide (BNP) for predicting all-cause mortality (C-statistic: z = 0.044, p = 0.965). These results suggest that BUN is an independent predictor of post-discharge all-cause mortality in elderly patients with ADHF and its prognostic performance was similar to that of BNP.


A Stochastic Binary Model for the Regulation of Gene Expression to Investigate Responses to Gene Therapy.
In this manuscript, we use an exactly solvable stochastic binary model for the regulation of gene expression to analyze the dynamics of response to a treatment aiming to modulate the number of transcripts of a master regulatory switching gene. The challenge is to combine multiple processes with different time scales to control the treatment response by a switching gene in an unavoidable noisy environment. To establish biologically relevant timescales for the parameters of the model, we select the RKIP gene and two non-specific drugs already known for changing RKIP levels in cancer cells. We demonstrate the usefulness of our method simulating three treatment scenarios aiming to reestablish RKIP gene expression dynamics toward a pre-cancerous state: (1) to increase the promoter's ON state duration; (2) to increase the mRNAs' synthesis rate; and (3) to increase both rates. We show that the pre-treatment kinetic rates of ON and OFF promoter switching speeds and mRNA synthesis and degradation will affect the heterogeneity and time for treatment response. Hence, we present a strategy for reaching increased average mRNA levels with diminished heterogeneity while reducing drug dosage by simultaneously targeting multiple kinetic rates that effectively represent the chemical processes underlying the regulation of gene expression. The decrease in heterogeneity of treatment response by a target gene helps to lower the chances of emergence of resistance. Our approach may be useful for inferring kinetic constants related to the expression of antimetastatic genes or oncogenes and for the design of multi-drug therapeutic strategies targeting the processes underpinning the expression of master regulatory genes.


Clinical trials in neuroprotection.
Neuroprotection is a therapeutic approach that aims to prevent neuronal degeneration and loss of function. Research has focused on developing neuroprotective agents for the therapy of various degenerative diseases, including Alzheimer's disease, amyotrophic lateral sclerosis, Parkinson's disease, and glaucoma. Clinical trials for the evaluation of neuroprotective agents pose unique challenges in terms of experimental design and data interpretation. In order to generate meaningful results, clinical trials on neuroprotective agents should ideally be designed to minimize the potential for bias and optimize the ability to detect the neuroprotective effect of a therapeutic intervention in as short a time as possible. Key issues for the design of clinical trials of neuroprotective therapies include identifying appropriate endpoints and determining the ideal timing of the intervention. Neuroprotection trials in glaucoma must be designed to distinguish between the neuroprotective effects of the therapy and the protective effect of intraocular pressure lowering. The choice of suitable functional endpoints in glaucoma trials is also a critical consideration. For example, visual field loss can be used as a functional endpoint; however, it occurs slowly and may require many years before meaningful changes occur. New methods for assessing visual function may be useful for assessing neuroprotective effects of therapeutic interventions. Although there have been a plethora of medications studied for neuroprotective effects in clinical trials, few have been approved by regulatory agencies for use in patients. Despite these challenges, properly designed clinical trials with validated endpoints will yield the most useful information on the neuroprotective effects of therapy, and may provide new treatment options to prevent the loss of neurologic function, including vision.


Knowledge, perceptions, and attitudes by residents in Punjab and Khyber Pakhtunkhwa, Pakistan in connection with bats.
Fruit bats play an important role in pollination and seed dispersal, and their conservation is important to maintain the productivity of some crops and natural ecosystems. The objective of this study was to investigate the knowledge, attitudes, and perception of fruit bats by orchard farmers and agricultural communities in Pakistan. The present survey was conducted in two districts (i.e. Sheikhupura and Malakand districts) within Punjab and Khyber Pakhtunkhwa provinces based on the higher number of fruit growing areas and bat roosting sites. A total of 200 (100 per district) close-ended questionnaires with 53 questions were administered to randomly selected respondents within the selected communities associated with fruit orchards, including orchard owners, laborers, and members of the surrounding community. Each questionnaire was divided into seven sections (i.e., demographic information, environmental and public health effects of bats, knowledge about bats, perception and control of bats, non-lethal methods adopted to control bats, and different myths about bats). A majority of respondents (59%, n = 118) mis-classified bats as birds instead of mammals despite more than 84% reporting that they have observed bats. Nearly 71.5% of orchard farmers perceived that their fruits are contaminated by bats during consumption, and a majority believe that bats destroy orchards (62.5%) and are responsible for spreading disease. Mythology about bats was ambiguous, as 49% of those surveyed did not perceived bats to bring good luck (49%), and 50% did not perceived them to be bad omens either. Most respondents have never killed a bat (68%) nor would they kill a bat if given the opportunity (95%). Regarding the control of bats, the greatest percentage of respondents strongly disagree with shooting bats (36%) and strongly agree with leaving bats alone (42.5%). This study provides a better understanding of the sociodemographic factors associated with knowledge, attitude and perception of bats from fruit orchard owners, labourers and local people. We recommend educational interventions for targeted groups in the community, highlighting the ecosystem services and importance of bat conservation to improve people's current knowledge regarding the role of bats and reduce direct persecution against bats.


Effects of Asp-369 and Arg-372 mutations on heme environment and function in human endothelial nitric-oxide synthase.
Eight polar amino acid residues in the putative substrate-binding region from Thr-360 to Val-379 in human endothelial nitric-oxide synthase (eNOS) (Thr-360, Arg-365, Cys-368, Asp-369, Arg-372, Tyr-373, Glu-377, and Asp-378) were individually mutated. Only two of these residues, Asp-369 and Arg-372, were found to be essential for enzyme activity. A further series of mutants was generated by replacing these two residues with various amino acids and the mutant proteins were expressed in a baculovirus system. Mutant eNOS had a very low L-citrulline formation activity with the exception of D369E and R372K, which retained 27% and 44% of the wild-type enzyme activity, respectively. Unlike the wild-type enzyme, all mutants except D369E, R372K, and R372M had a low spin heme (Soret peak at 416 nm). All the Asp-369 mutants had higher Kd values for L-arginine (1-10 mM) than wild-type eNOS (0.4 microM) and an unstable heme-CO complex, and except for D369E, had a very low (6R)-5,6,7, 8-tetrahydro-L-biopterin (BH4) content. In contrast, each of Arg-372 mutants retained a considerable amount of BH4, had a moderate reduction in L-arginine affinity, and had a more stable heme-CO complex. 1-Phenylimidazole did not bind to wild-type eNOS heme, but bound to all Asp-369 and Arg-372 mutants (Kd ranged from 10 to 65 microM) except R372K. Heme spin-state changes caused by binding of 3, 5-lutidine appeared to depend on both charge and size of the side chains of residues 369 and 372. Furthermore, all Asp-369 and Arg-372 mutants were defective in dimer formation. These results suggest that residues Asp-369 and Arg-372 in eNOS play a critical role in oxygenase domain active-site structure and activity.


The association between meniscal and cruciate ligament damage and knee pain in community residents.
To describe the frequency of meniscal and cruciate ligament damage by magnetic resonance imaging (MRI) and to examine its association with knee pain in community residents in Korea. Participants were randomly chosen regardless of knee osteoarthritis (OA) or pain from the population-based Hallym Aging Study. Demographic and knee pain data were obtained by questionnaire. Radiographic evaluations consisted of weight-bearing knee A-P radiographs and 1.5-T MRI scans. We assessed the integrities of the menisci and cruciate ligaments in the dominant knee of subjects without knee pain or in the more symptomatic knee among subjects with knee pain, and examined their association with knee pain using a logistic regression model. The mean age of the 358 study subjects was 71.8 years, and 51.4% were women. Meniscal and cruciate ligament damage were present in 49.7% and 8.0% of men and in 71.2% and 26.9% of women, respectively. The presence of meniscal damage was significantly associated with the presence of knee pain among subjects without radiographic knee OA (ROA), but not among subjects with ROA. The presence of cruciate ligament tear was associated with knee pain in subjects with or without ROA. The severity of knee pain was significantly correlated with medial meniscal damage grade but not with cruciate ligament tear. Incidental meniscal or cruciate findings on MRI were common in this elderly population. Among subjects without ROA, the presence of meniscal or cruciate damage was significantly associated with knee pain. The medial meniscal grade was significantly correlated with knee pain severity.


Natural variation of diterpenoid phytoalexins in rice: Aromatic diterpenoid phytoalexins in specific cultivars.
Rice (Oryza sativa L.) plants accumulate antimicrobial compounds known as phytoalexins in response to pathogen attack. To date, more than 20 compounds have been isolated as phytoalexins from rice, mostly diterpenoids. However, the quantitative analysis of diterpenoid phytoalexins in various cultivars has revealed that the cultivar 'Jinguoyin' does not accumulate these compounds at detectable concentrations. Therefore, in this study, we attempted to detect a new class of phytoalexins from Bipolaris oryzae infected leaves of 'Jinguoyin'. We detected five compounds in the leaves of the target cultivar, whereas these compounds were not detected in the leaves of 'Nipponbare' or 'Kasalath', which are representative cultivars of the japonica and indica subspecies. Subsequently, we isolated these compounds from ultraviolet (UV)-light-irradiated leaves and determined their structures by spectroscopic analysis and the crystalline sponge method. All the compounds were diterpenoids containing a benzene ring and were detected from the pathogen-infected rice leaves for the first time. Because the compounds showed antifungal activity against B. oryzae and Pyricularia oryzae, we propose that they function as phytoalexins in rice and named them abietoryzins A-E. The abietoryzins tended to accumulate at high concentrations in cultivars that accumulated low levels of known diterpenoid phytoalexins after UV-light irradiation. Of the total of 69 cultivars in the WRC, 30 cultivars accumulated at least one of the abietoryzins, and, in 15 cultivars, the amounts of some abietoryzins were the highest among those of the analyzed phytoalexins. Therefore, abietoryzins are a major phytoalexin group in rice, although their presence has, to date, been overlooked.


The tissue factor expression on CD14++CD16-monocytes is a new markers in the Chinese Han older adult population with sepsis: A prospective study.
To investigate the tissue factor (TF) expression on different subsets of monocyte and tissue factor secretion of peripheral blood and evaluate their association with the prognosis of sepsis in the Chinese older adult population. Monocyte subsets and TF expression on different subsets of monocyte were measured using flow cytometry in 80 older adult sepsis patients and 40 age and sex matched healthy controls. Plasma level of TF was measured using ELISA (enzyme-linked immunosorbent assay) method. TF expression on CD14++CD16- (MO1) monocyte was lower in death (28-day non-survivor) group compared with survival (28-day survivor) groups [1.01 % (0.58 %, 1.62 %) vs. 3.66 % (1.32 %, 6.93 %), p = 0.001]. The plasma level of TF was increased in death group compared to survival group according to the 28-day mortality [109.2 (67.3, 154.2) vs. 62.1 (44.7, 115.5) pg/mL, p = 0.031]. Logistic regression analysis showed TF expression on MO1 monocyte (β = -0.776, OR = 0.460, CI: 0.251, 0.843, p = 0.012) was independently associated with the 28-day mortality. The ROC (receiver operating characteristic) curve showed that the AUC (area under the curve) of the TF expression on MO1 monocyte for predicting 28-day mortality was 0.846 (p < 0.001). The TF expression on CD14++CD16- monocyte is a new marker for the prognosis of older adult sepsis.


Intra-seasonal decline of clutch size in Lesser Snow Geese.
Different mechanisms proposed to explain the intra-seasonal decline in clutch size of Lesser Snow Geese (Anser caerulescens caerulescens) were tested at the La Perouse Bay colony, Manitoba, Canada. Ovary examination of females collected after laying revealed that the actual number of eggs produced per female decreased over the laying period. This finding eliminates nest-parasitism, partial clutch predation and renesting attempts as sufficient explanations for seasonal clutch size decline. Follicular atresia induced by reserve depletion was also rejected since its occurrence was similar among early and late nesters. The decline in clutch size was observed within age-classes, and therefore age effects on clutch size and laying date per se do not account for the observed relationship. Clutch size and laying date were respectively positively and negatively correlated with the amount of nutrient reserves in females at the onset of laying, and also covaried within individuals observed breeding in several seasons. Laying date repeatability was estimated at 0.22. It is postulated that the seasonal decline in clutch size results mainly from a positive feed-back of female's nutrient reserves on the hormones controlling ovary development. Hatching synchrony or shortness of the breeding season may be the ultimate factors responsible for the intra-seasonal clutch size decline in Lesser Snow Geese.


Evaluation of a heat vulnerability index on abnormally hot days: an environmental public health tracking study.
Extreme hot weather conditions have been associated with increased morbidity and mortality, but risks are not evenly distributed throughout the population. Previously, a heat vulnerability index (HVI) was created to geographically locate populations with increased vulnerability to heat in metropolitan areas throughout the United States. We sought to determine whether areas with higher heat vulnerability, as characterized by the HVI, experienced higher rates of morbidity and mortality on abnormally hot days. We used Poisson regression to model the interaction of HVI and deviant days (days whose deviation of maximum temperature from the 30-year normal maximum temperature is at or above the 95th percentile) on hospitalization and mortality counts in five states participating in the Environmental Public Health Tracking Network for the years 2000 through 2007. The HVI was associated with higher hospitalization and mortality rates in all states on both normal days and deviant days. However, associations were significantly stronger (interaction p-value < 0.05) on deviant days for heat-related illness, acute renal failure, electrolyte imbalance, and nephritis in California, heat-related illness in Washington, all-cause mortality in New Mexico, and respiratory hospitalizations in Massachusetts. Our results suggest that the HVI may be a marker of health vulnerability in general, although it may indicate greater vulnerability to heat in some cases.


Magnetic nanoparticles of nitrogen enriched carbon (mnNEC) for analysis of pesticides and metabolites in zebrafish by gas chromatography-mass spectrometry.
Nanosized carbon based sorbents have been widely used for separation, enrichment and desalting of biological samples because of their distinguished characteristics. In this work, magnetic nanoparticles of nitrogen enriched carbon (mnNEC) have been developed for enrichment of organochlorine pesticide DDT and metabolite DDE that have been accumulated in zebrafish during the course of environmental exposure. Polymerization of pyrrole was performed in the aqueous suspension of Fe(3)O(4) nanoparticles. Resultant core-shell nanoparticules coated with polypyrrole were then subjected to a process of carbonization under high temperature and nitrogen atmospheric condition. The presence of nitrogen atoms in carbon nanoparticles increases the hydrophilicity and dispersability in aqueous samples. It has been experimentally demonstrated that mnNEC can be effectively dispersed in aqueous samples and rapidly isolated by the application of an external magnetic field. Recoveries of DDT and DDE from water range from 90% to 102% and 85-97% respectively. In combination with Selected Ion Monitoring (SIM) experiments of gas chromatography-mass spectrometry, the detection limit can be down to low ng/mL level. By using mnNEC approach, two interesting results have been found for zebrafish with 60 days exposure to DDT (1 μg/l). (1) There is higher concentration of DDT (37-143 ng/g) and DDE (173-1108 ng/g) in male zebrafish body tissues than that of female (7-52 ng/g and 146-362 ng/g for DDT and DDE respectively). (2) There is high ratio of DDE/DDT for both female and male zebrafish, implying high environmental persistence and ongoing bioaccumulation.


Clinical characteristics and prognosis of hospitalized COVID-19 patients with incident sustained tachyarrhythmias: A multicenter observational study.
Little is still known about the prognostic impact of incident arrhythmias in hospitalized patients with COVID-19. The aim of this study was to evaluate the incidence and predictors of sustained tachyarrhythmias in hospitalized patients with COVID-19, and their potential association with disease severity and in-hospital mortality. This was a retrospective multicenter observation study including consecutive patients with laboratory confirmed COVID-19 admitted to emergency department of ten Italian Hospitals from 15 February to 15 March 2020. The prevalence and the type of incident sustained arrhythmias have been collected. The correlation between the most prevalent arrhythmias and both baseline characteristics and the development of ARDS and in-hospital mortality has been evaluated. 414 hospitalized patients with COVID-19 (66.9 ± 15.0 years, 61.1% male) were included in the present study. During a median follow-up of 28 days (IQR: 12-45), the most frequent incident sustained arrhythmia was AF (N: 71; 17.1%), of which 50 (12.1%) were new-onset and 21 (5.1%) were recurrent, followed by VT (N: 14, 3.4%) and supraventricular arrhythmias (N: 5, 1.2%). Incident AF, both new-onset and recurrent, did not affect the risk of severe adverse events including ARDS and death during hospitalization; in contrast, incident VT significantly increased the risk of in-hospital mortality (RR: 2.55; P: .003). AF is the more frequent incident tachyarrhythmia; however, it not seems associated to ARDS development and death. On the other hand, incident VT is a not frequent but independent predictor of in-hospital mortality among hospitalized COVID-19 patients.


Modelling nonlinearity in piezoceramic transducers: From equations to nonlinear equivalent circuits.
Quadratic nonlinear equations of a piezoelectric element under the assumptions of 1D vibration and weak nonlinearity are derived by the perturbation theory. It is shown that the nonlinear response can be represented by controlled sources that are added to the classical hexapole used to model piezoelectric ultrasonic transducers. As a consequence, equivalent electrical circuits can be used to predict the nonlinear response of a transducer taking into account the acoustic loads on the rear and front faces. A generalisation of nonlinear equivalent electrical circuits to cases including passive layers and propagation media is then proposed. Experimental results, in terms of second harmonic generation, on a coupled resonator are compared to theoretical calculations from the proposed model.


Effect of Fat Grafting on Postoperative Edema After Orthognathic Surgery.
Autologous fat grafting concurrent with orthognathic surgery is a powerful adjunctive aesthetic tool, and has been shown to have anti-inflammatory properties in prior studies. The purpose of this study is to evaluate the effect of fat grafting on postoperative edema following orthognathic surgery, and also consider the impact of age and BMI. A retrospective cohort study was performed. Three-dimensional photos (Canfield, Fairfield, NJ) from a series of postprocedure time-points were analyzed using Geomagic Studio 2013 (3D Systems, Morrisville, NC). An unstructured covariance linear mixed model was created to analyze the effect and extent of the effect of age, BMI, and fat using SPSS Statistics 24.0 (IBM, Armonk, NY). One hundred thirty postoperative three-dimensional pictures were analyzed, from 31 patients. The linear mixed model demonstrates that the fat injected (P<0.001), age (P=0.001), and BMI (P<0.001) are significant factors in the postoperative volume. Age and BMI increase postoperative edema by 3.63 cm per year and 14.60 cm per kg/m, respectively. Fat injected reduces postoperative edema by 8.72 cm per 1 cc injected. Increasing age and BMI lead to greater postoperative edema (3.88 cm per year and 14.60 cm per kg/m, respectively). Fat grafting concurrent with orthognathic surgery reduces postoperative edema by 8.72 cm per 1 cc injected, and hastens return to steady state. This impact is more profound in patients with a greater age and BMI.


A case report of a novel compound heterozygous mutation in a Brazilian patient with deficiency of Interleukin-1 receptor antagonist (DIRA).
Deficiency of the natural antagonist of interleukin-1 was first described in 2009 and so far 20 patients has been reported. In Brazil just two cases have been reported both carrying the same homozygous 15 bp deletion. Blocking interleukin-1 has changed rate survival for DIRA patients. The use of anakinra and rilonacept has been reported safe and efficient, whereas the selective blockade of interleukin-1 beta, using the monoclonal antibody canakinumab has been reported in a single case only. Here we report a case of a 7 years old Brazilian boy that presented with recurrent episodes of systemic inflammation with severe disabling osteomyelitis with mild pustular skin rash. A Next Generation Sequencing gene panel allowed to detect two pathogenic mutations in the IL1RN gene, described in compound heterozygosity. Corticosteroids was effective in controlling inflammation and anti-IL1 beta blocker triggered disease flare. Complete clinical control could be achieved using IL-1 receptor antagonist. DIRA is a severe, life threatening autoinflammatory condition with low numbers of patients described all over the world. The mutation p.Asp72_Ile76del in IL1RN is presented in all Brazilian DIRA patients already described and p.Q45* (rs1019766125) is a new mutation affecting the IL1RN gene. Following the pathogenesis of DIRA, blocking both subunits of interleukin one as well as antagonizing the receptor using anakinra or rilonacept seems to be effective. There is just one report using canakinumab for the treatment of DIRA and this is the first report of disease flare using this drug.


Assessment of hypochondriasis.
This article gives a detailed description of the assessment of hypochondriasis. Other conditions are described in which this type of assessment can be useful. The aims of assessment are outlined, followed by a full description of the information that should be obtained, and methods of assessment used. Information gained at assessment is used to construct a cognitive-behavioural formulation of the problem, emphasising aetiological and maintaining factors, which then can be modified during treatment. Other important aspects of assessment include overcoming problems in the therapeutic relationship, and helping the patient to accept psychological treatment. Other practical difficulties in assessment are described, and some solutions are offered.


Intracochlea acoustic neuroma: a case report and discussion.
A case of an intracochlea acoustic neuroma that was diagnosed radiologically is presented and discussed.


Candida auris Blood stream infection- a descriptive study from Qatar.
Candida auris is an emerging yeast pathogen that can cause invasive infections, particularly candidemia, in healthcare settings. Candida auris is characterized by resistance to multiple classes of antifungal drugs and high mortality. To describe the risk factors, clinical characteristics, antifungal susceptibility pattern and outcomes of Candida auris blood stream infection. We conducted a retrospective review of electronic medical records of C. auris fungemia cases in the facilities under Hamad Medical corporation, Qatar from 1/11/2018 to 31/7/2021. Demographic data, risk factors, antibiogram and 30-day outcome are described. We identified 36 patients with C. auris fungemia. Most of the patients were in intensive care unit following severe COVID-19 pneumonia and had received steroids and broad-spectrum antibiotics. Most cases were central line related. Over 90% of isolates were non-susceptible to fluconazole, while amphotericin B resistance reached 85%. Factors associated with high mortality included initial SOFA score of 9 or above and absence of source control. Our study reveals a concerning 41.6% mortality rate within 30 days of C. auris candidemia. Furthermore, the prevalence of amphotericin B resistance in Qatar exceeds what has been reported in the literature necessitating further exploration. Echinocandins retains nearly 100% susceptibility and should be prioritized as the treatment of choice. These findings emphasize the need for vigilant monitoring and appropriate management strategies to combat C. auris infections and improve patient outcomes.


Cellular and humoral components of monocyte and neutrophil chemotaxis in cord blood.
Monocyte and polymorphonuclear neutrophil (PMN) chemotaxis was studied in cord blood from healthy term infants. Monocyte chemotaxis was normal to increased (115-126%) whereas PMN chemotaxis was decreased (79%) in comparison with that of healthy adult control subjects. Generation of chemotactic factors from cord sera was impaired, being 55% of that generated by pooled normal human serum (PNHS). Cord serum was less inhibitory than pooled adult human serum for adult monocytes when the cells were suspended in 10% serum and tested for chemotaxis. No inhibition of chemotactic factors by either cord or adult sera was observed. The dissociation of chemotactic response of the two different phagocytic cells may represent a protective mechanism whereby one cell can compensate for a defect in the response of the other.


NOX2 controls phagosomal pH to regulate antigen processing during crosspresentation by dendritic cells.
To initiate adaptative cytotoxic immune responses, proteolytic peptides derived from phagocytosed antigens are presented by dendritic cells (DCs) to CD8+ T lymphocytes through a process called antigen "crosspresentation." The partial degradation of antigens mediated by lysosomal proteases in an acidic environment must be tightly controlled to prevent destruction of potential peptides for T cell recognition. We now describe a specialization of the phagocytic pathway of DCs that allows a fine control of antigen processing. The NADPH oxidase NOX2 is recruited to the DC's early phagosomes and mediates the sustained production of low levels of reactive oxygen species, causing active and maintained alkalinization of the phagosomal lumen. DCs lacking NOX2 show enhanced phagosomal acidification and increased antigen degradation, resulting in impaired crosspresentation. Therefore, NOX2 plays a critical role in conferring DCs the ability to function as specialized phagocytes adapted to process antigens rather than kill pathogens.


Mechanical bowel preparation and antibiotic prophylaxis in colorectal surgery: use by and opinions of Spanish surgeons.
Antibiotic prophylaxis (AP) and mechanical bowel preparation (MBP) previous to surgery have classically been regarded as important in colorectal surgery. The latter has recently been questioned. We evaluated opinion of Spanish surgeons about the use of these measures. E-mail survey among all members of Spanish Coloproctologic Associations. Of 413 participants in the survey, 131 (31.7%) responded; 87% of surgeons used cathartics (70%), enemas (2%) or both (28%) for MBP. MBP was used 60% in right colon surgery, 90% in left colon and 99% in rectal surgery. Surgeons with more case load or those who specialized in colorectal surgery used significantly less MBP; 60% of the surgeons thought that MBP made surgery easier and reduced contamination; 35% thought that it decreased wound infection (WI) and 17% thought that it prevented anastomotic leaks. For 77%, it was regarded as useful or very useful. AP was used by 99.3% of surgeons including systemic alone in 86.2% and combined with oral in 16.8%. The first dose was given 2 h before surgery by 20.2% of the surgeons, at the anaesthetic induction by 78.3% and postoperatively by 1.5%; 43% used single dose only, 44.5% extended to 24 h and 12.5% for two or more days; 95% thought that AP reduced WI and 96% considered that it was useful. There is general agreement on AP. MBP remained a common practice among Spanish colorectal surgeons except for right colonic resection. Surgeons with more case load and specialization used it significantly less.


Mitochondrial rewiring drives metabolic adaptation to NAD(H) shortage in triple negative breast cancer cells.
Nicotinamide phosphoribosyltransferase (NAMPT) is a key metabolic enzyme in NAD+ synthesis pathways and is found upregulated in several tumors, depicting NAD(H) lowering agents, like the NAMPT inhibitor FK866, as an appealing approach for anticancer therapy. Like other small molecules, FK866 triggers chemoresistance, observed in several cancer cellular models, which can prevent its clinical application. The molecular mechanisms sustaining the acquired of resistance to FK866 were studied in a model of triple negative breast cancer (MDA-MB-231 parental - PAR), exposed to increasing concentrations of the small molecule (MDA-MB-231 resistant - RES). RES cells are not sensitive to verapamil or cyclosporin A, excluding a potential role of increased efflux pumps activity as a mechanism of resistance. Similarly, the silencing of the enzyme Nicotinamide Riboside Kinase 1 (NMRK1) in RES cells does not increase FK866 toxicity, excluding this pathway as a compensatory mechanism of NAD+ production. Instead, Seahorse metabolic analysis revealed an increased mitochondrial spare respiratory capacity in RES cells. These cells presented a higher mitochondrial mass compared to the FK866-sensitive counterparts, as well as an increased consumption of pyruvate and succinate for energy production. Interestingly, co-treatment of PAR cells with FK866 and the mitochondrial pyruvate carrier (MPC) inhibitors UK5099 or rosiglitazone, as well as with the transient silencing of MPC2 but not of MPC1, induces a FK866-resistant phenotype. Taken together, these results unravel novel mechanisms of cell plasticity to counteract FK866 toxicity, that, besides the previously described LDHA dependency, rely on mitochondrial rewiring at functional and energetic levels.


Upper limb sympathetic blockade following intercostal nerve blocks.
A case of transient sympathetic block of the arm following percutaneous intercostal nerve injections is described and the possible aetiology discussed.


Interaction of the electrophilic ketoprofenyl-glucuronide and ketoprofenyl-coenzyme A conjugates with cytosolic glutathione S-transferases.
Carboxylic acid-containing drugs are metabolized mainly through the formation of glucuronide and coenzyme A esters. These conjugates have been suspected to be responsible for the toxicity of several nonsteroidal anti-inflammatory drugs because of the reactivity of the electrophilic ester bond. In the present study we investigated the reactivity of ketoprofenyl-acylglucuronide (KPF-OG) and ketoprofenyl-acyl-coenzyme A (KPF-SCoA) toward cytosolic rat liver glutathione S-transferases (GST). We observed that KPF-SCoA, but not KPF-OG inhibited the conjugation of 1-chloro-2,4-dinitrobenzene and 4-nitroquinoline N-oxide catalyzed by both purified cytosolic rat liver GST and GST from FAO and H5-6 rat hepatoma cell lines. Photoaffinity labeling with KPF-SCoA suggested that the binding of this metabolite may overlap the binding site of 4-methylumbelliferone sulfate. Furthermore, high-performance liquid chromatography and mass spectrometry analysis showed that both hydrolysis and transacylation reactions were observed in the presence of GST and glutathione. The formation of ketoprofenyl-S-acyl-glutathione could be kinetically characterized (apparent K(m) = 196.0 +/- 70.6 microM). It is concluded that KPF-SCoA is both a GST inhibitor and a substrate of a GST-dependent transacylation reaction. The reactivity and inhibitory potency of thioester CoA derivatives toward GST may have potential implications on the reported in vivo toxicity of some carboxylic acid-containing drugs.


Edge-assisted quantum protocol for secure multiparty logical AND its applications.
Security and privacy have always been key concerns for individuals in various edge-assisted services. In this paper, we present a feasible quantum solution to an important primitive of secure multiparty computations, i.e., Secure Multiparty Logical AND (SMLA), in which n participants can securely compute logical AND of n private bits. In order to ensure perfect security and achieve good feasibility, we introduce a semi-honest edge server and two non-collusive fog nodes, and design a secure and feasible edge-assisted quantum protocol for SMLA, which cleverly utilizes Secure Multiparty XOR to implement SMLA group by group. Furthermore, we focus on applications of this quantum primitive protocol and design two quantum protocols for Multiple Private Set Intersection and Anonymous One-vote Veto. Compared with classical related protocols, our proposed quantum protocols obtain higher security, which can be guaranteed by the basic principles of quantum mechanics.


The influence of contextual factors on tic expression in Tourette's syndrome: a review.
Symptoms of Tourette's syndrome vary in frequency and intensity. Although such variability may be the result of deficits in the underlying neurological system, tic expression can also be systematically impacted by contextual factors. This article reviews research on the impact of several contextual factors on tic expression and discusses implications for future research and treatment development.


Systematic evaluation of the pre-eclampsia drugs, dietary supplements and biologicals pipeline using target product profiles.
The Accelerating Innovation for Mothers (AIM) project established a database of candidate medicines in research and development (R&D) between 2000 and 2021 for five pregnancy-related conditions, including pre-eclampsia. In parallel, we published target product profiles (TPPs) that describe optimal characteristics of medicines for use in preventing/treating pre-eclampsia. The study objective was to use systematic double screening and extraction to identify all candidate medicines being investigated for pre-eclampsia prevention/treatment and rank their potential based on the TPPs. Adis Insight, Pharmaprojects, WHO international clinical trials registry platform (ICTRP), PubMed and grant databases were searched (Jan-May 2021). The AIM database was screened for all candidates being investigated for pre-eclampsia. Candidates in clinical development were evaluated against nine prespecified criteria from TPPs identified as key for wide-scale implementation, and classified as high, medium or low potential based on matching to the TPPs. Preclinical candidates were categorised by product type, archetype and medicine subclass. The AIM database identified 153 candidates for pre-eclampsia. Of the 87 candidates in clinical development, seven were classified as high potential (prevention: esomeprazole, L-arginine, chloroquine, vitamin D and metformin; treatment: sulfasalazine and metformin) and eight as medium potential (prevention: probiotic lactobacilli, dalteparin, selenium and omega-3 fatty acid; treatment: sulforaphane, pravastatin, rosuvastatin and vitamin B3). Sixty-six candidates were in preclinical development, the most common being amino acid/peptides, siRNA-based medicines and polyphenols. This is a novel, evidence-informed approach to identifying promising candidates for pre-eclampsia prevention and treatment - a vital step in stimulating R&D of new medicines for pre-eclampsia suitable for real-world implementation.


Refractive index tuning of SiO2 for Long Range Surface Plasmon Resonance based biosensor.
A novel, highly sensitive and low cost Long Range Surface Plasmon Resonance (LRSPR) biosensor for detecting uric acid, as a model analyte, has been developed in this work. Silicon dioxide (SiO2) having low and tunable refractive index has been chosen as the dielectric layer for the excitation of LRSP modes replacing the most explored Cytop and Teflon polymers. The prepared LRSPR based uric acid bio-sensor gives good response characteristics with a high sensitivity of about 21.6°/mM and low limit of detection (LOD) of 0.02 mM. The fabricated LRSPR sensor was also evaluated to detect uric acid in real serum samples. The results yield a great scope to promote the development of robust, efficient and highly selective LRSPR based biosensors with SiO2 as tunable dielectric layer.


Solid-State Conformational Flexibility at Work: Zipping and Unzipping within a Cyclic Peptoid Single Crystal.
A peptidomimetic compound undergoes a reversible single-crystal-to-single-crystal transformation upon guest release/uptake with the transformation involving a drastic conformational change. The extensive and reversible alteration in the solid state is connected to the formation of an unprecedented "CH-π zipper" which can reversibly open and close (through the formation of CH-π interactions), thus allowing for guest sensing.


Motivation for physical activity before and after allogeneic hematopoietic stem cell transplantation.
This study aimed to identify patient motivation for physical activity before and after allogeneic hematopoietic stem cell transplantation (HSCT). We conducted 14 semi-structured interviews of seven patients (two of each patient): one before starting a conditioning regimen and one after leaving the protected environment. All interviews were recorded and analyzed using the inductive content analysis method. The data collection period was May to December 2018. The participants comprised three men and four women aged 40-70 years. The patients underwent bone marrow, umbilical cord blood, or peripheral HSCT. The patients' motivation for physical activity before and after HSCT was classified into six categories and categorized into five themes including "to overcome HSCT," "to look after myself," "to respond to the donor," "the existence of supporters," and "encouragement from supporters." The categories and themes developed here based on patient responses provide an important perspective that should be promoted among healthcare providers who care for patients undergoing HSCT.


Gone gene fishing: how to catch novel marine antimicrobials.
Medical or health-promoting products of marine origin are often regarded with skepticism--some, such as shark fins and cod liver oil, are frequently perceived as low-tech "alternative treatments" largely because they have not been exploited to their full potential. The marine environment is an enormous source of biodiversity--80% of all life is found under the oceans' surfaces--yet very little of this rich resource has been utilized. Furthermore, most marine organisms rely heavily on antimicrobial components of their innate immune defenses to combat pathogens. The past three years has seen a revolution in the methods used to identify novel antimicrobials from marine sources; among the most promising are marine cationic antimicrobial peptides (CAPs).


Instantaneous Formation of Silk Protein Aerosols and Fibers with a Portable Spray Device Under Ambient Conditions.
A variety of artificial silk spinning approaches have been attempted to mimic the natural spinning process found in silkworms and spiders, yet instantaneous silk fiber formation with hierarchical structure under physiological and ambient conditions without post-treatment procedures remains unaddressed. Here, we report a new strategy to fabricate silk protein-based aerosols and silk fibers instantaneously (< 1 s) in situ using a simple, portable, spray device, avoiding complicated and costly advanced manufacturing techniques. The key to success is the instantaneous conformational transition of silk fibroin from random coil to β-sheet right before spraying by mixing silk and polyethylene glycol (PEG) solutions in the spray device, allowing aerosols and silk fibers to be sprayed in situ, with further control achieved via the molecular weight of silk. The spinning process of the spray device is based on the use of green solvents, i.e., all steps of instant conformational transition of silk fibroin are carried out in aqueous conditions or with buffers at ambient conditions, in combination with shear and elongational flow caused by the hydraulic pressure generated in the spray container. The system supports a portable and user-friendly system that could be used for drug delivery carriers, wound coating materials and rapid silk fiber conformal coatings on surfaces.


The clinical value of texture analysis of dual-time-point 18F-FDG-PET/CT imaging to differentiate between 18F-FDG-avid benign and malignant pulmonary lesions.
To examine whether the texture analysis of dual-time-point (DTP) F-18-fluorodeoxyglucose (18F-FDG)-PET/CT imaging can differentiate between 18F-FDG-avid benign and malignant pulmonary lesions. We compared standardized uptake value (SUV)-related (SUVmax [g/ml] and SUVmean [g/ml]), volumetric (metabolic tumor volume [MTV] [cm3] and total lesion glycolysis [TLG] [g]), and texture (entropy, homogeneity, dissimilarity, intensity variability [IV], size-zone variability [SZV], and zone percentage [ZP]) (MTV ≥ 5.0 cm3 and SUV ≥ 2.5 g/ml) parameters between 13 benign and 46 malignant lesions using the Mann-Whitney U test. Diagnostic performance was evaluated by receiver operating characteristic (ROC) analysis. Stepwise logistic regression analysis was performed to identify and use the independent variables that correctly differentiate between benign and malignant lesions. Malignant pulmonary lesions showed significantly higher SUVmax, SUVmean, MTV, TLG, entropy, dissimilarity, IV, and SZV and significantly lower homogeneity and ZP than benign pulmonary lesions (all p < 0.05) in both early and delayed images. Their areas under the ROC curves (AUCs) ranged between 0.69 and 0.94, and diagnostic accuracies between 64.4% and 93.2%. Entropy-early (p = 0.014), SUVmean-delay (p = 0.039), and dissimilarity-delay (p = 0.027) were independent parameters, and combined use of them yielded the highest AUC (0.98) with 100% sensitivity (46/46), 84.6% specificity (11/13), and 96.7% (57/59) accuracy for distinguishing between benign and malignant lesions. The individual early and delayed SUV-related, volumetric, and texture parameters showed a wide range of accuracy. Combined use of independent parameters extracted from DTP imaging might yield a high diagnostic accuracy with balanced sensitivity and specificity to differentiate between benign and malignant 18F-FDG-avid pulmonary lesions. • Malignant pulmonary lesions showed significantly higher SUV-related (SUVmax and SUVmean) and volumetric (MTV and TLG) parameters than benign pulmonary lesions in both early and delayed images. • Malignant pulmonary lesions showed significantly more heterogeneous18F-FDG uptake than benign pulmonary lesions in both early and delayed images. • Combined use of independent parameters extracted from DTP imaging might yield a high diagnostic accuracy to differentiate between benign and malignant18F-FDG-avid pulmonary lesions.


Semantic processing in children with Cochlear Implants: A review of current N400 studies and recommendations for future research.
Deaf and hard of hearing children with cochlear implants (CI) often display impaired spoken language skills. While a large number of studies investigated brain responses to sounds in this population, relatively few focused on semantic processing. Here we summarize and discuss findings in four studies of the N400, a cortical response that reflects semantic processing, in children with CI. A study with auditory target stimuli found N400 effects at delayed latencies at 12 months after implantation, but at 18 and 24 months after implantation effects had typical latencies. In studies with visual target stimuli N400 effects were larger than or similar to controls in children with CI, despite lower semantic abilities. We propose that in children with CI, the observed large N400 effect reflects a stronger reliance on top-down predictions, relative to bottom-up language processing. Recent behavioral studies of children and adults with CI suggest that top-down processing is a common compensatory strategy, but with distinct limitations such as being effortful. A majority of the studies have small sample sizes (N < 20), and only responses to image targets were studied repeatedly in similar paradigms. This precludes strong conclusions. We give suggestions for future research and ways to overcome the scarcity of participants, including extending research to children with conventional hearing aids, an understudied group.


Urban-rural disparities in the prevalence and trends of depressive symptoms among Chinese elderly and their associated factors.
This study aimed to examine urban-rural disparities in the prevalence and trends of depressive symptoms (DS) among Chinese elderly and associated factors. A total of 8025, 7808, and 4887 respondents aged 60 years and above were selected from the China Family Panel Studies (CFPS) in 2016, 2018, and 2020, respectively. DS was assessed using a short version of Center for Epidemiologic Studies Depression Scale (CES-D). Twenty-two associated factors from six categories were included in random forest (RF) models. All urban-rural comparisons were conducted based on good model performance. The DS prevalence among all rural elderly was significantly higher than corresponding urban elderly. This disparity continued to widen among younger elderly, while it continued to narrow among older elderly. The top 10 common leading factors were sleep quality, self-rated health, life satisfaction, memory ability, child relationship, IADL disability, marital status, educational level, and gender. Urban-rural disparities in sleep quality, interpersonal trust, and child relationship continued to widen, while disparities in multimorbidity, hospitalization status, and frequency of family dinner continued to narrow. This study may exist recall bias and lacks causal explanation. Significant and continuing disparities in the DS prevalence were observed between urban and rural elderly in China, showing opposite trends in younger and older elderly. The top 10 leading associated factors for DS were nearly consistent across urban and rural elderly, with sleep quality having strongest contribution. Urban-rural disparities in associated factors also showed different trends. This study provides a reference for mental health promotion among Chinese elderly.


Comparison among three different fixation techniques in temporal brow lift surgery.
The eyebrow lifting with temporal approach is an increasing widespread technique. Many surgical procedures are described, but the long-term stability is dependent on the effectiveness in the fixation of elevated tissues. The authors compared 3 different fixation techniques in temporal brow lift. Forty-five consecutive patients aged between 33 and 70 underwent brow lift surgery with temporal approach. Patients were divided into 3 groups; in group 1, anchorage was performed with Endotine Ribbon, group 2 with a Mersilene mesh, and group 3 with a Prolene suture. The amount of brow elevation was assessed comparing the distance between interpupillar line and superior eyebrow hairline, measured at the midpupil and at the lateral and medial canthal angle. The follow-up was 1 year. All patients had a pleasant improvement in brow shape. The average initial brow position was 19.84 mm near the head of the eyebrow area, 20.74 mm at the body, and 19.57 mm in the region of the tail. A progressive recovery and a partial relapse regarding the eyebrow body (23.88 mm at 6 months and 23.02 mm at 1 year) occurred, but overall for the lateral region, it passed from 27.53 mm at 6 months to 25.80 mm after a 12-month follow-up. Final brow position was dependent upon surgical technique used in fixation. Different options in brow elevation and stabilization affect the final shape and position of the brows. From the statistical analysis, mesh suspension provided the best results in terms of long-lasting stability. Therapeutic III.


[Experiences with endoscopic placed endoprosthesis in stenosing malignancies of the esophagus and cardia].
Seventy-five patients with obstructing esophagogastric malignancies were palliatively treated by fiberendoscopic insertion of prosthetic tubes. The indications for endoscopic intubation are discussed, the procedure is described. The mortality rate was 6.6%. Complications were obstructions due to food impaction (10), tumor overgrowth (5) or mucosa folds (3), reflux and aspiration (7), dislocation of the prosthesis (5), perforation (1) and desintegration of the tube (1). Normal swallowing was restored in 98% and in general, there was a marked improvement in the quality of life. In summary, we conclude palliative fiberendoscopic intubation as the best existing procedure in patients with an incurable obstructing esophagogastric neoplasm.


Identification and characterization of proteins that interact with Drosophila melanogaster protein kinase CK2.
D. melanogaster CK2 (DmCK2) is a highly conserved protein kinase that is composed of catalytic, alpha, and regulatory, beta, subunits associated as an alpha2beta2 heterotetramer. In order to analyze the functions of CK2 in this metazoan model, we have used the two hybrid approach to identify interacting proteins. One of these cDNAs, DmA24, encodes a novel polypeptide with no homologs in GenBank, and is notable in that it contains a bipartite nuclear localization signal and two sites for phosphorylation by CK2. In situ hybridization to polytene chromosomes indicates that the DmA24 gene is located at the 61 D interval of chromosome II a region that also harbors 3 additional genes with similar structure. DmA24p interacts with DmCK2alpha, but not with DmCK2beta, demonstrating that this interaction is specific for the catalytic subunit of CK2. In addition, the protein is phosphorylated by the holoenzyme purified from Drosophila embryos. These studies identify DmA24p as a potentially new physiological partner of DmCK2. In addition, we also report the results of a large-scale screen that has identified a new set of DmCK2-interacting proteins. Most notable among these are Surf6, a nucleolar protein involved in RNA processing, and Spalt, a homeotic protein.


Cervical osteomyelitis after placement of a self-expanding plastic stent for palliation of dysphagia associated with chemoradiation-induced esophageal strictures.
Esophageal strictures are a common sequela of chemoradiation and/or surgery to the head and neck cancers and can lead to stenosis and significant dysphagia. Endoscopic dilation endoscopic and placement of self-expanding stents are often to used relieve dysphagia symptoms. However, these stents are not without risks and complications. We present a case of a 58-year-old man who had the rare complication of cervical osteomyelitis as a result of plastic esophageal stent placement for palliation of chemoradiation-induced strictures. The patient was successfully managed with immobilization of the cervical spine in a halo vest and appropriate antibiotics. To the best of our knowledge, this is the first reported case of cervical spine osteomyelitis after self-expanding plastic stent (SEPS) placement for esophageal stricture. It was successfully treated with immobilization and antibiotic therapy. The treating physician should be aware of this rare complication to make an early diagnosis. Literature on esophageal stent-induced cervical osteomyelitis is reviewed.


CD71 surface analysis of T cells: a simple alternative for extracorporeal photopheresis quality control.
Extracorporeal photopheresis (ECP) is a leukapheresis-based cellular therapy that is used with increasing frequency worldwide to treat various T-cell-mediated diseases. Currently, the inhibition of T-cell proliferation after photopheresis is analysed frequently using time-consuming assays including radioactive thymidine assays or carboxyfluorescein succinimidyl ester (CFSE) staining. We investigated whether simple surface T-cell staining using surrogate markers of T-cell proliferation can replace time-consuming measurement of T-cell proliferation in ECP quality control. T-cell activation markers were investigated by flow cytometry after ECP. Candidates were validated by direct comparison with the classical CFSE T-cell proliferation inhibition test and apoptosis staining. Finally, surface T-cell staining was performed in patient samples in comparison with classical methods. CD71 expression exhibited the fastest and most robust upregulation, which was detectable as early as 6-8 h after T-cell stimulation and almost completely abrogated by ECP. In a direct comparison with the CFSE T-cell proliferation assay, suppression of CD71 expression after ECP was almost identical and detectable as early as 16 h after stimulation in peripheral blood mononuclear cells of healthy donors. Furthermore, in direct comparison with classical apoptosis staining, the inhibition delta of CD71 after ECP was significantly higher. Moreover, in patients under T-cell suppressive therapy, T-cell-dependent CFSE and CD71 assays exhibited decreased sensitivity to detect ECP treatment and were inferior in comparison to apoptosis staining. Surface CD71 analysis represents a very simple quality control alternative to detect ECP-mediated T-cell proliferation inhibition in normal PBMC samples devoid of T-cell suppressive drugs.


The hallmarks of ovarian cancer: proliferation and cell growth.
Epithelial ovarian cancer (EOC) is a heterogeneous group of diseases with distinct biological and clinical behaviour. Despite the differences between them, the capability of tumour cells to continuously proliferate and avoid death is maintained among histotypes. This ability is the result of alterations at different levels, causing the deregulation of cell cycle and proliferative-related pathways. Even if the leading role is played by RB and TP53, changes in other molecular pathways are involved in the development of EOC. This ability can be exploited to generate in vitro and in vivo models resembling the conditions of tumour development in a patient. In vivo models, such as patient-derived xenografts (PDX) or genetically engineered mouse models (GEMM), represent a fundamental tool in the study of the molecular mechanisms implicated in each EOC biotype for testing new therapeutic approaches. Herein we describe the major proliferation-related pathways and its disruption found in EOC and how these features can be used to establish in vivo models for translational research.


Clinical impact of timely reporting of IgM for West Nile Virus.
In 2002 diagnostic testing for West Nile Virus (WNV) infection was not ideal because commercial products were unavailable. Although testing was performed without charge by public health (PH), testing is not intended to be used for patient management and generally results are not available in a timely or reliable manner. This study was conducted to determine the clinical impact of having WNV serology available by a diagnostic laboratory within a few days of sample collection. An IgM immunoassay (FT, from Focus Technologies) was performed August 13, 2003 to October 31, 2003 Monday, Wednesday and Friday in our hospital's virology laboratory with split sample testing at PH. All results (positive and negative) were called to physicians. Turn-around time for the FT assay was within 0-3 days; for PH, it was approximately 8 days. Of 95 samples, 6 were positive and 89 negative. For the FT negative samples, all were concordant with PH, but four FT positive samples (three sera and one cerebrospinal fluid) were discrepant. After resolution of the discrepancies, the FT test had a sensitivity of 100% and a specificity of 99%. Physicians reacted positively to having results faster. Fifty-eight percent of the results had clinical impact, 22% had undetermined impact, 13% were patient requests, and 4% had no impact. Concluding, the FT assay is 100% sensitive and 99% specific and in 58% there was clinical impact for WNV reporting in a timely manner.


Integrating multi-omics data to investigate pseudogene expression in Mycolicibacterium smegmatis.
Large-scale -omics data now allows for investigating genome-wide functional elements. Using RNA-Seq data, we tested the expression of 134 pseudogenes which do not show duplicated protein coding genes in the M. smegmatis genome. We observe significant expression and translation of 28 pseudogenes. Further examination using RNA-Seq reads suggested the sequencing errors in many pseudogenes. These include some of the functionally relevant genes such as recN and manB. We propose that the analysis of transcriptional and translational landscape using multi-dimensional -omics data could shed light on the current annotations of the bacterial pseudogenes.


Effects of incentives for naltrexone adherence on opiate abstinence in heroin-dependent adults.
To test whether an incentive-based intervention that increased adherence to naltrexone also increased opiate abstinence. Post-hoc combined analysis of three earlier randomized controlled trials that showed individually that incentives for adherence to oral and to extended-release injection naltrexone dosing schedules increased naltrexone adherence, but not opiate abstinence. Out-patient therapeutic work-place in Baltimore, MD, USA. One hundred and forty unemployed heroin-dependent adults participating from 2006 to 2010. Participants were hired in a model work-place for 26 weeks and randomized to a contingency (n = 72) or prescription (n = 68) group. Both groups were offered naltrexone. Contingency participants were required to take scheduled doses of naltrexone in order to work and earn wages. Prescription participants could earn wages independent of naltrexone adherence. Thrice-weekly and monthly urine samples tested for opiates and cocaine and measures of naltrexone adherence (percentage of monthly urine samples positive for naltrexone or percentage of scheduled injections received). All analyses included pre-randomization attendance, opiate use and cocaine use as covariates. Additional analyses controlled for cocaine use and naltrexone adherence during the intervention. Contingency participants had more opiate abstinence than prescription participants (68.1 versus 52.9% opiate-negative thrice-weekly urine samples, respectively; and 71.9 versus 61.7% opiate-negative monthly urine samples, respectively) based on initial analyses [thrice-weekly samples, odds ratio (OR) = 3.3, 95% confidence interval (CI) = 1.7-6.5, P < 0.01; monthly samples, OR = 2.6, 95% CI = 1.0-7.1, P = 0.06] and on analyses that controlled for cocaine use (thrice-weekly samples, OR = 3.9, 95% CI = 3.3-4.5, P < 0.01; monthly samples, OR = 3.4, 95% CI = 1.1-11.1, P = 0.04), which was high and associated with opiate use. The difference in opiate abstinence rates between contingency and prescription participants was reduced when controlling for naltrexone adherence (monthly samples, OR = 1.1, 95% CI = 0.7-1.7, P = 0.84). Incentives for naltrexone adherence increase opiate abstinence in heroin-dependent adults, an effect that appears to be due to increased naltrexone adherence produced by the incentives.


Identification of key genes for diabetic kidney disease using biological informatics methods.
Diabetic kidney disease (DKD) is a common complication of diabetes, which is characterized by albuminuria, impaired glomerular filtration rate or a combination of the two. The aim of the present study was to identify the potential key genes involved in DKD progression and to subsequently investigate the underlying mechanism involved in DKD development. The array data of GSE30528 including 9 DKD and 13 control samples was downloaded from the Gene Expression Omnibus database. The differentially expressed genes (DEGs) in DKD glomerular and tubular kidney biopsy tissues were compared with normal tissues, and were analyzed using the limma package. Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed for DEGs using the GO Function software in Bioconductor. The protein‑protein interaction (PPI) network was then constructed using Cytoscape software. A total of 426 genes (115 up‑ and 311 downregulated) were differentially expressed between the DKD and normal tissue samples. The PPI network was constructed with 184 nodes and 335 edges. Vascular endothelial growth factor A (VEGFA), α‑actinin‑4 (ACTN4), proto‑oncogene, Src family tyrosine kinase (FYN), collagen, type 1, α2 (COL1A2) and insulin‑like growth factor 1 (IGF1) were hub proteins. Major histocompatibility complex, class II, DP α1 (HLA‑DPA1) was the common gene enriched in the rheumatoid arthritis and systemic lupus erythematosus pathways, and the immune response was a GO term enriched in module A. VEGFA, ACTN4, FYN, COL1A2, IGF1 and HLA‑DPA1 may be potential key genes associated with the progression of DKD, and immune mechanisms may serve a part in DKD development.


The Healthy LifeWorks project: The effect of a comprehensive workplace wellness program on the prevalence and severity of musculoskeletal disorders in a Canadian government department.
The purpose of this study was to determine the effect of a comprehensive workplace wellness program on the prevalence and severity of musculoskeletal disorders in a Canadian government department. The Healthy LifeWorks program was developed, implemented, and evaluated over a 4-year period. A total of 233 employees completed the Nordic Musculoskeletal Questionnaire before and after the program to determine the prevalence and severity of musculoskeletal disorders. There was an approximately 10% decrease in the 12-month prevalence of musculoskeletal disorders, ranging from 4% for hip/thigh problems to 12% for lower and upper back problems. The proportion of people reporting that a musculoskeletal disorder interfered with their normal work during the past 12 months decreased from 83% to 46%. Comprehensive wellness, including educational sessions on posture, ergonomics, and joint health, results in improved musculoskeletal health.


From cadaveric and animal studies to the clinical reality of robotic mastectomy: a feasibility report of training program.
Robotic nipple-sparing mastectomy (RNSM) provides better cosmetic outcomes and improves the quality of life of women with breast cancer. However, this has not been widely adapted due to the lack of well-structured training programs. The present study aimed to report the establishment of cadaveric and animal skill laboratory training programs for RNSM and the participants' perception on the training programs. We performed 24 RNSMs using 11 cadavers and one porcine model. Then, the skill laboratory characteristics were reviewed. Five trainers and 10 trainees participated in the programs. The first four cadaveric RNSMs with latissimus dorsi flaps and implants were performed using the da Vinci Si® system. We performed 14 and six RNSMs using the Xi® and SP® systems, respectively. The scores for questionnaires on the satisfaction with the training consisted of the trainees' perceived goals in attending the course, teaching/learning environment, and teaching staff performance. The scores were excellent. Cadaveric or porcine RNSM skill laboratory training may be essential programs that can provide safe and efficient training.


Immunogenic properties of rapidly digested food proteins following gavage exposure of mice: a comparison of ovalbumin with a potato acid phosphatase preparation.
The ability of food proteins to resist digestion in simulated gastric fluid (SGF) correlates with allergenic potential. The purpose of the current investigations was to determine whether this association is due solely to the failure of unstable proteins to elicit an immune response when administered orally. We have examined immune responses induced in BALB/c mice by gavage administration of ovalbumin (OVA) and a crude potato protein extract (PPE) containing acid phosphatase activity. The stability of OVA and PPE in SGF was measured using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The ability of these proteins to stimulate specific IgG and IgE antibody production in mice following parenteral (intraperitoneal; ip) or oral (gavage) exposure was compared using enzyme-linked immunosorbent and homologous passive cutaneous anaphylaxis assays, respectively. Both OVA and PPE induced specific IgG antibody responses when administered either by gavage or by ip injection. Parenteral, but not gavage, exposure to OVA was associated with robust IgE antibody responses. Administration of PPE failed to stimulate strong IgE production via either route of exposure. Differential stability in SGF was observed, with PPE being digested extremely rapidly (within 1 min), whereas OVA was more resistant. The strong association reported by others between stability in SGF and allergenic potential is unlikely to be solely due to orally-ingested labile proteins failing to provoke immune responses due to degradation in the stomach.


Data on the role of the frontal cortex in vegetative behavior.
A homeostatic conditional reflex (CR) was elaborated to the effect of repeated inhalations in rats of a gas mixture containing 8 percent oxygen. The effect of the conditional response was opposite to that of the unconditional one. After ablation of the frontal cortex, the conditional reaction disappeared. The repeated administration of 40 mg/kg of histamine resulted in a tolerance to the temperature lowering effect of histamine similar to the habituation. An injection of distilled water brought about dishabituation. Tolerance was not influenced by the ablation of the frontal cortex, but the dishabituating effect of distilled water was absent.


Undesirable effects of citrus juice on the pharmacokinetics of drugs: focus on recent studies.
It is well known that intake of grapefruit juice affects the pharmacokinetics of various kinds of drugs. It has been reported that other citrus juices also interact with certain drugs. To re-evaluate citrus juice-drug interactions based on currently available evidence, a literature search was conducted for new and updated information since the grapefruit juice-drug interaction was last reviewed in 1998. MEDLINE (1998-October 2004) was accessed and more than 200 reports were found. The effects of grapefruit juice ingestion on the pharmacokinetics of orally administered drugs have been reported for 40 drugs since the reviews published in 1998. Increases in either area under the concentration-time curve (AUC) or maximum plasma concentration (C(max)) were found with 34 of these, the major mechanism being considered to be inactivation of intestinal cytochrome P450 3A4, a so-called mechanism-based inhibition. Although recent reports point to the inhibitory effects of grapefruit juice on the function of P-glycoprotein, which transports substrates from enterocytes back into the lumen, the contribution to the bioavailability of drugs that are substrates of P-glycoprotein has not been established yet. Dramatic decreases in AUC and C(max) for two drugs in association with grapefruit juice ingestion has been reported and, in these cases, inhibitory effects on organic anion transporting polypeptide, which mediates absorption from the intestinal lumen to enterocytes, might be involved. Other citrus juices such as Seville (sour) orange juice and commonly ingested varieties of orange juice also showed significant effects on the AUC and C(max) of some drugs. Although the situation is complex and uncertainties remain, we recommend that patients avoid citrus juice intake while taking medications and that healthcare providers advise against citrus juice intake in this setting until any interactions with subject drugs can be clarified in clinical studies.


Biological activity of saponins and saponin-like compounds from starfish and brittle-stars.
Twenty-four saponins and saponin-like compounds, isolated from starfish and brittle-stars, have been tested in four in vitro tests, based upon bacterial and cell tissue cultures. Saponin-like compounds from brittle-stars have previously not been tested for biological activity. In an antibacterial test based on an agar diffusion test, the Gram positive bacterium S. aureus was affected by the polyhydroxylated steroidal glycosides, polyhydroxylated sterols and disulfated sterols. However, none of the 21 compounds tested were active against the Gram negative bacterium E. coli. In a cytotoxicity test all 21 compounds tested influenced the cells at a concentration of 100 micrograms/ml, while the cells were unaffected at 1 microgram/ml. In an antitumor test, 16 compounds were tested on two lymphoma cell lines. Inhibition of cell growth, at a concentration of 5 ng/ml, was seen for three polyhydroxylated sterols, in one cell line. Weak activity was seen in an antiviral test at a concentration of 10 micrograms/ml.


Evaluation of the usability of two types of image display systems, during laparoscopy.
This study was performed to assess the optimal display location of a flat-screen monitor for laparoscopy. It was also performed to assess the posture (objective), opinion, and preference (subjective) of subjects using a flat-screen monitor positioned in the optimal display location and a cathode-ray tube monitor on a tower next to the operating table (current situation). Twelve surgeons performed cholecystectomies using the two display systems alternately. The postures of the operator and the assistant were assessed by an infrared video analysis system. The posture of the assistant is significantly better when using a flat-screen monitor [more neutral head flexions (p = 0.036) and neutral neck torsions (p = 0.012)]. No significant differences were found for the posture of the operator. The operators and assistants felt more comfortable when using a flat-screen monitor (p = 0.008) and they preferred this display to the use of a monitor on a tower. The use of flat-screen monitors is better for the physical and psychological comfort of the users, even though the technical performance is inferior in comparison with that of regular monitors.


Range extension of Lepidocephalichthys alkaia (Teleostei: Cobitidae) and notes on its sexual dimorphism.
The natural distributional range of the cobitid loach Lepidocephalichthys alkaia is extended into Yunnan Province, China. The modified sexually dimorphic pectoral fin in males of L. alkaia is described.


The relative importance of sexual reproduction versus clonal spread in an aridland bunchgrass.
Festuca idahoensis (Idaho fescue) is a perennial caespitose grass, common in semi-arid rangelands of the Intermountain West. To determine how individuals are recruited into a population, we studied two long-term monitoring plots that were established in 1937 at the Northern Great Basin Experimental Range in southeastern Oregon. The plots measured 3.05x3.05 m, and were located approximately 30 m apart. One plot was ungrazed, the other was subject to moderate levels of cattle grazing. The number of F. idahoensis plants in both plots increased ten-fold between 1937 and 1996, but whether this was due primarily to reproduction by seed or clonal fragmentation was unknown. In 1996, we mapped and sampled 160 plants of F. idahoensis. We used dominant inter-simple sequence repeat (ISSR) markers and codominant allozyme markers in order to identify genetic individuals and measure genetic diversity. Both plots were characterized by high levels of genetic and clonal diversity. When information from ISSRs, allozymes and sample location were combined, 126 genets were recognized, each consisting of one to four samples (ramets). By measuring the diameter of clones surrounding plants that were present in 1937, we estimated that clonal spread occurred at a rate of approximately 3.7 cm per decade, and thus was of secondary importance in the maintenance and increase of F. idahoensis stands. Sexual reproduction, rather than clonal fragmentation, accounted for most of the recruitment of new plants into these plots. The grazed plot had fewer ramets, genotypes, and clones than the ungrazed plot, but the ramets were significantly larger. Levels of genetic diversity did not differ in the grazed and ungrazed plots, but there was some evidence for a small, but significant level of genetic differentiation between the two. The results also indicate that F. idahoensis has the potential to be a long-lived species with some individuals persisting in excess of 60 years. This study demonstrates how long-term monitoring can be supplemented by genetic analysis to obtain detailed information on the population dynamics of plants. In the case of this community dominant species, this provides essential information for understanding succession and developing management and restoration strategies.


Perceptions of malaria transmission, presentation and management in northern Cameroon.
During early 2001, 1197 adult health service users in a poor rural district of northern Cameroon were interviewed in order to discover prevalent beliefs about malaria. The survey included questions about the name of the disease, its cause and transmission, signs and symptoms, cure, and local importance as a cause of sickness and death. Interviewees showed good understanding of the signs and symptoms of malaria, its importance, and its association with the rains. Most felt that either a medical or a herbal treatment could cure malaria. Only 1% identified mosquitoes as a source of transmission. Health education to explore beliefs and to correct misconceptions should precede and accompany the introduction of technological interventions in malaria control programmes in traditional societies.


Study of the effect of vitamin D supplementation on glycemic control in type 2 diabetic prevalent hemodialysis patients.
Vitamin D is claimed to have an adjuvant effect on glycemic control by dual action on pancreatic β-cells and insulin resistance. The aim of this study was to assess the possible effect of short-term alfacalcidol supply on glycemic control in type 2 diabetic hemodialysis (HD) patients. Twenty type 2 diabetic HD patients (using diet and oral drugs but not insulin) were randomly selected from our dialysis unit as well as 20 non-diabetic HD patients as control. A third group of 12 healthy subjects were studied as well. All three groups were similar in age, sex, and body mass index. Oral alfacalcidol therapy was administrated daily as recommended by Kidney Disease Outcomes Quality Initiative (K/DOQI) guidelines for 12 weeks guided by monthly serum phosphorus and Cax PO4 product. Corrected total calcium, phosphorus, intact parathyroid hormone, 25-hydroxy vitamin D (25[OH]D), and glucoparameters (fasting blood glucose, glycated hemoglobin [HbA1c%], insulin resistance by homeostatic model assessment, and β-cell function by HOMA-β%) were measured under basal conditions and after 3 months of therapy. 25(OH)D was non-significantly lower in diabetic than non-diabetic HD patients, but significantly lower than healthy subjects at the start of the study. However, vitamin D level increased significantly after 3 months of trial, although the levels did not reach normal values. This vitamin D rise was associated with highly significant improvement in concentrations of fasting blood sugar (FBS), fasting insulin, HbA1c%, and HOMA-β-cell function in diabetic and non-diabetic controls. However, there was a significant rise in insulin resistance after treatment. The percentage of change was evident more in diabetics regarding FBS and 25(OH)D concentration. Adjustment of 25(OH)D level in type 2 diabetic prevalent HD patients may improve, at least with short-term therapy, glycemic control mainly through improving β-cell function.


Use of the lactose H2 breath test to monitor mucosal healing in coeliac disease.
Confirmation of the diagnosis of coeliac disease requires unequivocal evidence of recovery on a gluten-free diet. The lactose H2 breath test is a non-invasive technique for detecting lactose malabsorption, and this may occur in untreated coeliac disease. The utility of this test was investigated in objectively confirming a response to gluten exclusion. The study included 44 children from 0.9 to 14.75 y of age (median 3.2) with coeliac disease. Five children were asymptomatic, being identified through coeliac antibody screening. Breath tests were performed prior to dietary treatment. If positive, they were repeated at 4-wk intervals following gluten exclusion. Overall, 21/44 (48%) children had positive breath tests at presentation. In 18/21 children on a gluten-free diet, this became negative after 4 wk (86%) and in all the children by 8 wk. In two children with positive tests at 4 wk, problems with dietary adherence were identified. A positive test before treatment was not associated with significant differences in individual symptoms, height, weight and body mass index standard deviation scores, serum haemoglobin or albumin. However, no positive results were found in the asymptomatic patients (p = 0.05). Lactose malabsorption was associated with earlier age of presentation (p = 0.008). The lactose H2 breath test objectively confirms a response to gluten exclusion. In selected cases it can help confirm a diagnosis of coeliac disease. However, the test tends to be less informative in asymptomatic patients, and in older children.


Contamination status of hexachlorobenzene (HCB) in sandstorm depositions from Beijing and the origination region.
In China, sandstorms play an important role in the long-range atmospheric transport and deposition of hexachlorobenzene (HCB). Concentrations of HCB in sandstorm depositions sampled from Beijing and the origination region were measured to reveal HCB levels during atmospheric transport. The results suggested a constant level of HCB during atmospheric transport. The values were close to environmental background values indicating that no potential source of HCB existed along the transport route. A tendency for HCB levels in Beijing to increase from north to east was also observed, which was probably due to the influence of wind speed and industrial zone distribution.


How being female influenced my professional experiences and growth.
In this paper I describe some of my professional experiences as a female, both as a graduate student and throughout my career. My own experience was unique because I began graduate school a few months after arriving in the U.S. with limited knowledge of English in a very competitive and demanding program , in addition to the fact that I was a faculty wife in that department. A major dilemma in achieving a successful academic career involves finding an optimal balance between the demands of a career and those of motherhood. In earlier years the burden of compromises dictated by the needs of children customarily fell on the shoulders of women, and this often delayed the development of their careers. This has changed some over the years as many couples share child-rearing responsibilities, but remains a hurdle for many professional women. While institutional biases and conflicting demands still produce special challenges to women who strive to make significant contributions to their fields, such women are more in charge of their careers today than they were 30 or 40years ago. But in my experience, the achievement of contributing significantly to knowledge in the field, for both men and for women, involves quite a few sacrifices.


Icariin-loaded 3D-printed porous Ti6Al4V reconstruction rods for the treatment of necrotic femoral heads.
Avascular necrosis of the femoral head is a prevalent hip joint disease. Due to the damage and destruction of the blood supply of the femoral head, the ischemic necrosis of bone cells and bone marrow leads to the structural changes and the collapse of the femoral head. In this study, an icariin-loaded 3D-printed porous Ti6Al4V reconstruction rod (referred to as reconstruction rod) was prepared by 3D printing technology. The mechanical validity of the reconstruction rod was verified by finite element analysis. Through infilling of mercapto hyaluronic acid hydrogel containing icariin into the porous structure, the loading of icariin was achieved. The biological efficacy of the reconstruction rod was confirmed through in vitro cell experiments, which demonstrated its ability to enhance MC3T3-E1 cell proliferation and facilitate cellular adhesion and spreading. The therapeutic efficacy of the reconstruction rod was validated in vivo through a femoral head necrosis model using animal experiments. The results demonstrated that the reconstruction rod facilitated osteogenesis and neovascularization, leading to effective osseointegration between bone and implant. This study provides innovative strategy for the treatment of early avascular necrosis of the femoral head. STATEMENT OF SIGNIFICANCE: The bioactivity of medical titanium alloy implants plays an important role in bone tissue engineering. This study proposed a medicine and device integrated designed porous Ti6Al4V reconstruction rod for avascular necrosis of the femoral head, whose macroscopic structure was customized by selective laser melting. The bionic porous structure of the reconstruction rod promoted the growth of bone tissue and formed an effective interface integration. Meanwhile, the loaded icariin promoted new bone and vascular regeneration, and increased the bone mass and bone density. Therefore, the implantation of reconstruction rod interfered with the further development of necrosis and provided a positive therapeutic effect. This study provides innovative strategies for the treatment of early avascular necrosis of femoral head.


Clinical features and magnetic resonance imaging characteristics of diskospondylitis in dogs: 23 cases (1997-2010).
To describe the signalment, clinical features, and most common MRI characteristics in dogs with diskospondylitis and investigate whether a correlation exists between the degree of spinal cord compression and neurologic status of the patient. Retrospective case series. 23 dogs. The medical records and imaging database of the Veterinary Teaching Hospital at Washington State University were retrospectively cross-referenced for cases of diskospondylitis in dogs from 1997 through 2010. Signalment, clinical signs, MRI characteristics, and results of bacteriologic cultures of urine, blood, CSF, or intervertebral disk material were reviewed. On T2-weighted sequences, vertebral endplates were most often of mixed signal intensity, whereas the vertebral body was hypointense. The intervertebral disk space was most often hyperintense on T2-weighted and short tau inversion recovery sequences and of mixed signal intensity on T1-weighted sequences. Paravertebral soft tissue hyperintensities were noted commonly on T2-weighted and short tau inversion recovery sequences. Heterogenous contrast enhancement of endplates and intervertebral disk spaces also occurred commonly, whereas contrast enhancement of vertebral bodies and paravertebral soft tissues was uncommon. Intramedullary spinal cord intensity was noted at 10 of 27 sites on T2-weighted sequences. Static spinal cord compression occurred in 17 of 23 dogs, and a significant direct correlation was found between the percentage of spinal cord compression and the patient neurologic score. Results suggested that diskospondylitis in dogs has a characteristic MRI appearance, and in some patients, MRI may aid in the identification of severe spinal cord compression, which could warrant surgical intervention.


Chromium occurrence and speciation in Baltimore harbor sediments and porewater, Baltimore, Maryland, USA.
Industrial activities in the Baltimore Harbor, Baltimore, Maryland, USA, have resulted in widespread chromium contamination of sediments. A comprehensive analysis of Cr speciation in sediment and porewater collected from 22 locations in the Baltimore Harbor was completed to understand Cr bioavailability and probability of toxicity due to Cr in sediments. The analysis employed a reverse-phase ion-pair high-performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) method. Sub-microgram-per-liter determination of Cr(III) and Cr(VI) in environmental samples was found, with method validation revealing broad method applicability of HPLC-ICP-MS to a wide range of sample types. The major limitation of the method was poor Cr species separation in high ionic strength solutions (greater than 0.1 M NaCl). Total Cr concentrations in Baltimore Harbor sediments ranged from 2.5 to 1,050 mg/kg with 11 of the 22 sites containing total Cr in excess of the 370 mg/ kg effects range-median (ER-M) sediment quality guideline. The Cr(VI) concentrations in sediments, however, were markedly lower, ranging from 0.10 to 0.38 mg/kg with Cr(VI) not detected in 14 of the 22 stations. Porewater concentrations, both for total Cr and Cr(VI), were quite low, with total Cr ranging from 0.20 to 2.16 microg/L and Cr(VI) ranging from 0.73 to 1.17 microg/L. The Cr(VI)-reducing capacity of the sediments, based on a sediment-spiking experiment, was found to be strongly correlated with the acid volatile sulfides content of the sediment. Overall, our results provide field validation of the hypothesis that Cr(VI) will not persist in sediments with excess acid volatile sulfides. Given the low concentrations of Cr(VI) in sediment and porewater, it appears unlikely that Cr in Baltimore Harbor sediments contributes appreciably to previously observed sediment toxicity.


Lipoprotein lipase in heart cell cultures is suppressed by bacterial lipopolysaccharide: an effect mediated by production of tumor necrosis factor.
Exposure of rat heart cell cultures, consisting mainly of nonbeating mesenchymal cells, to 50 ng/ml of bacterial lipopolysaccharide (LPS) for 24 h resulted in a more than 80% reduction in lipoprotein lipase activity. The loss of enzymic activity was accompanied by a concomitant reduction in enzyme protein, as shown by immunoblotting. Addition of LPS to the culture medium resulted also in the production of tumor necrosis factor (TNF), and the fall in lipoprotein lipase in LPS-treated cultures could be prevented by an antibody to TNF. Addition of recombinant human TNF to the heart cell cultures also depressed lipoprotein lipase activity. LPS treatment of preadipocytes in culture resulted in a fall in lipoprotein lipase activity and TNF production. Since TNF is known as a macrophage product, the cultures were tested for phagocytic capacity, and only 0.2-1.3% of the cells were shown to engulf Staphylococcus albus. Immunofluorescent staining with monoclonal antibodies OX-1, which identify leukocyte common antigen, was negative, and only 0.1 +/- 0.07% of the cells were positive after staining with OX-42 antibody to iC3b receptor. Both antibodies stained more than 98% of rat peritoneal macrophages used as controls. Since LPS treatment of macrophages at numbers comparable to or exceeding the number of phagocytic cells present in the heart cell cultures did not induce measurable amounts of TNF, it is suggested that in the heart cell cultures, TNF may be produced by cells other than macrophages.


Modeling the role of endoplasmic reticulum-mitochondria microdomains in calcium dynamics.
Upon inositol trisphosphate (IP3) stimulation of non-excitable cells, including vascular endothelial cells, calcium (Ca2+) shuttling between the endoplasmic reticulum (ER) and mitochondria, facilitated by complexes called Mitochondria-Associated ER Membranes (MAMs), is known to play an important role in the occurrence of cytosolic Ca2+ concentration ([Ca2+]Cyt) oscillations. A mathematical compartmental closed-cell model of Ca2+ dynamics was developed that accounts for ER-mitochondria Ca2+ microdomains as the µd compartment (besides the cytosol, ER and mitochondria), Ca2+ influx to/efflux from each compartment and Ca2+ buffering. Varying the distribution of functional receptors in MAMs vs. the rest of ER/mitochondrial membranes, a parameter called the channel connectivity coefficient (to the µd), allowed for generation of [Ca2+]Cytoscillations driven by distinct mechanisms at various levels of IP3 stimulation. Oscillations could be initiated by the transient opening of IP3 receptors facing either the cytosol or the µd, and subsequent refilling of the respective compartment by Ca2+ efflux from the ER and/or the mitochondria. Only under conditions where the µd became the oscillation-driving compartment, silencing the Mitochondrial Ca2+ Uniporter led to oscillation inhibition. Thus, the model predicts that alternative mechanisms can yield [Ca2+]Cyt oscillations in non-excitable cells, and, under certain conditions, the ER-mitochondria µd can play a regulatory role.


c-Jun N-terminal kinase (JNK) signaling contributes to cystic burden in polycystic kidney disease.
Polycystic kidney disease is an inherited degenerative disease in which the uriniferous tubules are replaced by expanding fluid-filled cysts that ultimately destroy organ function. Autosomal dominant polycystic kidney disease (ADPKD) is the most common form, afflicting approximately 1 in 1,000 people. It primarily is caused by mutations in the transmembrane proteins polycystin-1 (Pkd1) and polycystin-2 (Pkd2). The most proximal effects of Pkd mutations leading to cyst formation are not known, but pro-proliferative signaling must be involved for the tubule epithelial cells to increase in number over time. The c-Jun N-terminal kinase (JNK) pathway promotes proliferation and is activated in acute and chronic kidney diseases. Using a mouse model of cystic kidney disease caused by Pkd2 loss, we observe JNK activation in cystic kidneys and observe increased nuclear phospho c-Jun in cystic epithelium. Genetic removal of Jnk1 and Jnk2 suppresses the nuclear accumulation of phospho c-Jun, reduces proliferation and reduces the severity of cystic disease. While Jnk1 and Jnk2 are thought to have largely overlapping functions, we find that Jnk1 loss is nearly as effective as the double loss of Jnk1 and Jnk2. Jnk pathway inhibitors are in development for neurodegeneration, cancer, and fibrotic diseases. Our work suggests that the JNK pathway should be explored as a therapeutic target for ADPKD.


High Frequency of Juxtamembrane Domain ERBB2 Mutation in Gastric Cancer.
ERBB2 mutation is an emerging therapeutic target in solid tumors; its therapeutic responses depend on the location of mutation. In gastric cancer, the profiles of ERBB2 mutations and their relationship with human epidermal growth factor receptor 2 (HER2) overexpression remain unknown. We aimed to describe the details of ERBB2 mutations in gastric cancer. Comprehensive panel sequencing was performed in 234 advanced gastric cancer patients. We investigated hotspots and clinicopathologic features of ERBB2 mutant gastric cancer in a single institute and evaluated the hotspots of ERBB2 mutation in a public database. Eighteen patients (7.7%) had ERBB2 mutations. The most frequent mutation was p.Arg678Gln (42.1%), which was located in the juxtamembrane domain and was the most common mutation in public databases (20.5%). All 18 ERBB2-mutant patients were negative for HER2 expression. Co-occurring genetic alterations included KRAS, PIK3CA, and ATM mutations. ERBB2 mutations were not associated with HER2 overexpression in gastric cancer patients. The most common mutation was located in the juxtamembrane domain of ERBB2.


The population dynamics of nitrifiers in ammonium-rich systems.
Non-optimal pH, dissolved oxygen concentration, the presence of toxic substances, or the influence of grazers are known to cause disturbances in nitrification. Because activated sludge is a mixture of different organisms, bacteria, and higher organisms, the stability of processes such as carbon removal, nitrification, denitrification, and dephosphatation depends on a range of interactions. These interactions occur both between and within trophic levels. Understanding of the ecology of microorganisms involved in bioprocesses is essential for effective control of startup and operation of a particular process. The aim of the study was to gain further insight into the dynamics of nitrifiers in activated sludge at various sludge ages while treating higher concentrations of ammonium. The results confirmed the importance of Nitrosococcus mobilis and Nitrobacter sp. as the dominant nitrifiers responsible for nitritation and nitratation, respectively, in the presence of unlimited ammonium. The size of the dominant bacteria colony was larger compared to the other species present and reached 25 microm. Problems with nitrification occurred in all high-ammonium loaded reactors. The dynamics of nitrifier population was monitored by oxygen uptake rate (OUR) using a test enabling the OUR measurement separately for ammonium-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). The results reveal the hypersensitivity of nitrifiers to the substrate and products of incomplete nitrification.