Review: the impacts of circulating 25-hydroxyvitamin D levels on cancer patient outcomes: a systematic review and meta-analysis.
Circulating levels of 25-hydroxyvitamin D [25(OH)D] may affect the prognosis of cancer patients; however, the epidemiological results are not consistent. To perform a meta-analysis of all published studies to assess the associations of circulating 25(OH)D levels measured at or near the time of diagnosis and outcomes for cancer patients. Searches of the PubMed and MEDLINE databases were performed and updated to December 2013. Studies reporting an association between circulating 25(OH)D levels at or near the time of diagnosis and outcomes for the patients were included. Data extraction was performed independently by two authors, and conflicts were resolved by a third investigator. Included in the meta-analysis were 25 studies with 17 332 cases. Significant associations between circulating 25(OH)D levels at or near the time of diagnosis and the outcomes for cancer patients were found. The pooled hazard ratio for the highest vs the lowest quartile of circulating 25(OH)D levels was 0.55 (95% confidence interval [CI] = 0.33-0.91) for overall survival of colorectal cancer patients, 0.63 (95% CI = 0.51-0.77) for breast cancer patients, and 0.48 (95% CI = 0.36-0.64) for lymphoma patients. Higher 25(OH)D levels were significantly associated with reduced cancer-specific mortality for patients with colorectal cancer (P = .005) and lymphoma (P < .001) and improved disease-free survival for patients with breast cancer (P < .001) or lymphoma (P < .05). A 10-nmol/L increment in circulating 25(OH)D levels conferred a hazard ratio of 0.96 (95% CI = 0.95-0.97) for overall survival of the cancer patients. The results indicate that cancer patients with higher circulating 25(OH)D levels at or near the time of diagnosis have better outcomes.


Detection of delayed hypersensitivity to Fonsecaea pedrosoi metabolic antigen (chromomycin).
An experimental study was conducted between January 2002 and April 2003 for the detection of delayed hypersensitivity to Fonsecaea pedrosoi metabolic antigen (chromomycin) in skin tests. A total of 194 subjects were attended by spontaneous demand at the Infectious and Parasitic Diseases outpatient clinic of the Federal University of Maranhão-UFMA and at the Department of Microbiology, Federal University of Minas Gerais-UFMG and classified into three groups: patients with chromoblastomycosis caused by F. pedrosoi (n=20), healthy subjects (n=86) and patients with other diseases (n=88). For the skin test, 0.1 ml of the antigen was applied to the anterior side of the right forearm and 0.1 ml Smith medium was applied to the anterior side of the left forearm as control. The results were analyzed 48 h after inoculation of the antigen and an induration >/= 5 mm was considered to indicate a positive test. A cellular immune response to chromomycin was detected in 18 (90.0%) of the 20 patients with chromoblastomycosis caused by F. pedrosoi, and one of the patients with a negative test had reactional leprosy. Eighty-five (98.8%) of the 86 healthy subjects presented a negative reaction and only one reacted positively to the antigen. The skin test was negative in all 88 (100%) patients with other diseases, such as dermatophytosis, paracoccidioidomycosis, pulmonary aspergilloma, candidiasis, pityriasis versicolor, tuberculosis, leprosy, tegumentary leishmaniasis and syphilis, and one case of chromoblastomycosis caused by Rhinocladiella aquaspersa. Chromomycin was effective in detecting delayed hypersensitivity in patients with chromoblastomycosis caused by F. pedrosoi, with a sensitivity and specificity of 90.0% and 98.8%, respectively. These results suggest that this antigen can be used in the auxiliary diagnosis of the disease and also in epidemiological studies for determination of the prevalence of chromoblastomycosis infection in endemic areas.


Impact of timing of feed delivery on the behavior and productivity of dairy cows.
The objective of this work was to assess the effect of timing of feed delivery on the behavior and productivity of cows milked 3 times per day. Twelve lactating Holstein dairy cows (4 primiparous and 8 multiparous), milked 3 times per day (at 1400, 2100, and 0700 h), were individually assigned and exposed to each of 2 treatments (over 21-d periods) in a replicated crossover design. Treatments were the manipulation of timing of TMR delivery, 2 times per day, in relation to milking time: (1) feeding at milking time (at 1400 and 0700 h), and (2) feeding halfway between milking times (at 1730 and 1030 h). Milk production, feeding, sorting, and rumination behavior were monitored for each animal for the last 7 d of each treatment period. Milk samples were collected for 2 of the last 4 d of each period for milk component analysis. With a feed delivery delay, dry matter intake (DMI) tended to be lower (26.5 vs. 27.2 kg/d). Although no difference was found in feeding time (224.2 min/d), cows fed with a delay consumed their feed more slowly (0.12 vs. 0.13 kg of dry matter/min) in more frequent meals (10.0 vs. 9.1 meals/d), which were smaller in size (2.8 vs. 3.1 kg/meal) and tended to be shorter in duration (26.7 vs. 30.1 min/meal). Cows fed at milking sorted for long particles (102.3%), whereas cows fed with a delay did not sort for or against those particles. Cows sorted for medium particles to a similar extent (102.5%) on each treatment. Cows did not sort for or against short particles on either treatment. Sorting against fine particles was observed, to a similar extent (97.1%), on both treatments. Rumination time (8.9 h/d) and lying time (9.5 h/d) were similar between treatments. Cows without fresh feed at the 1400 h milking tended to stand for less time following that milking (71.0 vs. 94.0 min), whereas cows without fresh feed at the 0700 h milking stood for less time following that milking (66.3 vs. 87.8 min). No difference in this latency to lie down was seen at the 2100 h milking. Milk yield (48.0 kg/d), milk fat content (3.64%), and milk protein content (2.86%) were similar between treatments. Given the tendency for a difference in DMI and no change in yield, efficiency of production was improved with a feed delay (1.93 vs. 1.80 kg of milk/kg of DMI). These data suggest that moving the timing of feed delivery resulted in cows consuming their feed more slowly in smaller, more frequent meals, contributing to an improvement in efficiency of production.


Mutations in the slow skeletal muscle fiber myosin heavy chain gene (MYH7) cause laing early-onset distal myopathy (MPD1).
We previously linked Laing-type early-onset autosomal dominant distal myopathy (MPD1) to a 22-cM region of chromosome 14. One candidate gene in the region, MYH7, which is mutated in cardiomyopathy and myosin storage myopathy, codes for the myosin heavy chain of type I skeletal muscle fibers and cardiac ventricles. We have identified five novel heterozygous mutations--Arg1500Pro, Lys1617del, Ala1663Pro, Leu1706Pro, and Lys1729del in exons 32, 34, 35, and 36 of MYH7--in six families with early-onset distal myopathy. All five mutations are predicted, by in silico analysis, to locally disrupt the ability of the myosin tail to form the coiled coil, which is its normal structure. These findings demonstrate that heterozygous mutations toward the 3' end of MYH7 cause Laing-type early-onset distal myopathy. MYH7 is the fourth distal-myopathy gene to have been identified.


Bayesian data integration and variable selection for pan-cancer survival prediction using protein expression data.
Accurate prognostic prediction using molecular information is a challenging area of research, which is essential to develop precision medicine. In this paper, we develop translational models to identify major actionable proteins that are associated with clinical outcomes, like the survival time of patients. There are considerable statistical and computational challenges due to the large dimension of the problems. Furthermore, data are available for different tumor types; hence data integration for various tumors is desirable. Having censored survival outcomes escalates one more level of complexity in the inferential procedure. We develop Bayesian hierarchical survival models, which accommodate all the challenges mentioned here. We use the hierarchical Bayesian accelerated failure time model for survival regression. Furthermore, we assume sparse horseshoe prior distribution for the regression coefficients to identify the major proteomic drivers. We borrow strength across tumor groups by introducing a correlation structure among the prior distributions. The proposed methods have been used to analyze data from the recently curated "The Cancer Proteome Atlas" (TCPA), which contains reverse-phase protein arrays-based high-quality protein expression data as well as detailed clinical annotation, including survival times. Our simulation and the TCPA data analysis illustrate the efficacy of the proposed integrative model, which links different tumors with the correlated prior structures.


Effects of implant design on marginal bone changes around early loaded, chemically modified, sandblasted Acid-etched-surfaced implants: a histologic analysis in dogs.
A minimal marginal bone loss around implants during early healing has been considered acceptable. However, the preservation of the marginal bone is related to soft tissue stability and esthetics. Implant designs and surfaces were evaluated to determine their impact on the behavior of the crestal bone. The purpose of this study is to evaluate histologic marginal bone level changes around early loaded, chemically modified, sandblasted acid-etched-surfaced implants with a machined collar (MC) or no MC (NMC). Three months after a tooth extraction, 72 sandblasted acid-etched chemically modified implants were placed in six dogs. Thirty-six implants had NMC, and 36 implants had a 2.8-mm MC. All implants were loaded 21 days after placement. For histologic analyses, specimens were obtained at 3 and 12 months. Assessments of the percentage of the total bone-to-implant contact and linear measurements of the distance from the shoulder of the implant to the first bone-to-implant contact (fBIC) were performed. Based on fBIC measurements, estimates of bone loss were obtained for each implant. A mixed-model analysis of variance was used to assess the effects of implant type and sacrifice time. All implants achieved osseointegration. The mean bone gain observed around NMC early loaded implants (at 3 months: 0.13 ± 0.37 mm; at 12 months: 0.13 ± 0.44 mm) was significantly different from the mean bone loss for MC early loaded implants (at 3 months: -0.32 ± 0.70 mm; at 12 months: -0.79 ± 0.35 mm) at 3 months (P = 0.003) and 12 months (P <0.001). No infrabony component was present at the marginal fBIC around NMC implants in most cases. There were no statistically significant differences among the means of total bone contact for implant types. Chemically modified, sandblasted acid-etched-surfaced implants with NMC presented crestal bone gain after 3 and 12 months under loading conditions in the canine mandible. The implant design and surface were determinants in the marginal bone level preservation.


The many different prices paid to providers and the flawed theory of cost shifting: is it time for a more rational all-payer system?
In developed nations that rely on multiple, competing health insurers-for example, Switzerland and Germany-the prices for health care services and products are subject to uniform price schedules that are either set by government or negotiated on a regional basis between associations of health insurers and associations of providers of health care. In the United States, some states-notably Maryland-have used such all-payer systems for hospitals only. Elsewhere in the United States, prices are negotiated between individual payers and providers. This situation has resulted in an opaque system in which payers with market power force weaker payers to cover disproportionate shares of providers' fixed costs-a phenomenon sometimes termed cost shifting-or providers simply succeed in charging higher prices when they can. In this article I propose that this price-discriminatory system be replaced over time by an all-payer system as a means to better control costs and ensure equitable payment.


Signature of quantum Griffiths singularity state in a layered quasi-one-dimensional superconductor.
Quantum Griffiths singularity was theoretically proposed to interpret the phenomenon of divergent dynamical exponent in quantum phase transitions. It has been discovered experimentally in three-dimensional (3D) magnetic metal systems and two-dimensional (2D) superconductors. But, whether this state exists in lower dimensional systems remains elusive. Here, we report the signature of quantum Griffiths singularity state in quasi-one-dimensional (1D) Ta2PdS5 nanowires. The superconducting critical field shows a strong anisotropic behavior and a violation of the Pauli limit in a parallel magnetic field configuration. Current-voltage measurements exhibit hysteresis loops and a series of multiple voltage steps in transition to the normal state, indicating a quasi-1D nature of the superconductivity. Surprisingly, the nanowire undergoes a superconductor-metal transition when the magnetic field increases. Upon approaching the zero-temperature quantum critical point, the system uncovers the signature of the quantum Griffiths singularity state arising from enhanced quenched disorders, where the dynamical critical exponent becomes diverging rather than being constant.


Use of modern birth control methods among rural communities in Imo State, Nigeria.
This paper studied the extent of utilization of Modern Birth Control Methods (MBCM) among rural dwellers in Imo State Nigeria. Three hundred and sixty households were randomly selected and data were obtained from them with the use of questionnaires and Focus Group Discussion. The results showed that only 30% of the respondents used MBCM while 57% of them used the traditional birth control methods. The most popular modern method was the condom (24.2%). This was followed by the IUD, used by only 2.5% of the respondents. Some of the identified factors that hindered the use of MBCM included perceived negative health reaction, fear of the unknown effects, cost, spouse's disapproval, religious belief and inadequate information. For a better understanding and utilization of MBCM, it is recommended that adequate educational campaign should be mounted in the rural areas on the advantages of MBCM. This campaign should target the men the more because they make the major decisions on MBCM use.


Clinical Reasoning: Longitudinally Extensive Spinal Cord Lesions in a Middle-aged Man.
An immunocompetent 47-year-old man presented with a five-month history of progressive lower limb weakness, back pain, sphincter dysfunction, and intermittent fever, suggesting myelopathy in a chronic deteriorating course. A comprehensive analysis comprising of blood tests, neuroimaging, CSF profiling, molecular analysis, and histopathology was performed. Notably, enhanced spinal cord MRI revealed longitudinally extensive intradural-extramedullary lesions involving the cervical, thoracic, and lumbosacral spinal cord, with homogeneous enhancement and spinal cord compression. Serum treponema pallidumhemagglutination (TPHA) and rapid plasma reagin (RPR) tests were positive. CSF profiling showed pleocytosis, significant protein elevations, hypoglycorrhachia, and positive TPHA test. 18F-FDG-PET/CT indicated slightly increased intraspinal fluorodeoxyglucose (FDG) uptake. Spinal cord biopsy further showed small round blue cells in poorly differentiated tissues. Immunostaining was positive for NKX2.2, CD56, CD99, Synaptophysin, and Ki67 (50%). Molecular analysis detected a novel MALAT-CYSLTR1 fusion protein and variants in oncogenic genes including PTCH1, TERT, CREBBP, SPEN, and STK11 The diagnosis of intraspinal extraosseous Ewing sarcoma (ES) was confirmed. Briefly, our case details the diagnosis of a patient with intradural-extramedullary ES and highlights the value of spinal cord biopsy in progressive myelopathy of unknown causes.


Potent and selective partial ecdysone agonist activity of chromafenozide in Sf9 cells.
Chromafenozide (ANS-118) is a non-steroidal ecdysone mimic and its insecticidal effect is highly specific to lepidoptera. In order to evaluate the transcription-inducing activity via nuclear ecdysone receptor (EcR) and the mode of action of chromafenozide, ecdysone-responsive reporter gene assay systems were developed in Sf9 and Kc cells. Ponasterone A, a full EcR agonist, induced reporter transcription in a dose-dependent manner in both Sf9 and Kc cells. In contrast, chromafenozide activated reporter transcription with comparable potency to ponasterone A only in Sf9 cells, although its maximum activity was 4-fold lower than that of ponasterone A. When chromafenozide was applied together with ponasterone A to Sf9 cells, it antagonized ponasterone A at nanomolar concentrations. These results suggest that chromafenozide is a potent partial EcR agonist specific to lepidoptera; it appears to bind lepidopteran EcR with comparable affinity to ponasterone A, but may activate the EcR in a different manner.


Prostate-Specific Antigen Trends Predict the Probability of Prostate Cancer in a Very Large U.S. Veterans Affairs Cohort.
If prostate-specific antigen (PSA) trends help identify elevated prostate cancer (PCa) risk, they might provide early warning of progressing cancer for further evaluation and justify annual testing. Our objective was to determine whether PSA trends predict PCa likelihood. A biopsy cohort of 361,657 men was obtained from a Veterans Affairs database (1999-2012). PSA trends were estimated for the 310,458 men with at least 2 PSA tests prior to biopsy. Cancer tumors may grow exponentially with cells doubling periodically. We hypothesized that PSA from prostate cancer grows exponentially above a no cancer baseline. We estimated PSA trends on that basis along with five descriptive variables: last PSA before biopsy, growth rate in PSA from cancer above a baseline, PSA variability around the trend, number of PSA tests, and time span of tests. PSA variability is a new variable that measures percentage deviations of PSA tests from estimated trends with 0% variability for a smoothly increasing trend. Logistic regression models were used to estimate relationships between the probability of PCa at biopsy and the trend variables and age. All five PSA trend variables and age were significant predictors of prostate cancer at biopsy (p < 0.0001). An overall logistic regression model achieved an AUC of 0.67 for men with at least 4 tests over at least 3 years, which was a substantial improvement over a single PSA (AUC 0.58). High probability of PCa was associated with low PSA variability (smooth trends), high PSA, high growth rate, many tests over a long time-span and older age. For example, at 4.0 PSA the probability of cancer is 32% for 1 PSA test and increases to 68% for 8 tests over 7 years with smooth, fast growth (0% variability and 50% exponential growth). Our results show that smooth, fast exponential growth in PSA above a baseline predicts an increased probability of PCa. The probability increases as smooth (low variability) trends are observed for more tests over a longer time span, which makes annual testing worth considering. Worrisome PSA trends might be used to trigger further evaluation and continued monitoring of the trends-even at low PSA levels.


Dos and Don'ts in Pregnancy: Truths and Myths.
Pregnancy is a time of excitement and anxiety. The reality for pregnant women is that their actions could affect their pregnancies and their fetuses. As such, they need to know what they should and should not do to minimize risk and optimize outcomes. Whereas this advice used to come from doctors, a few books, and some family and friends, in the age of the internet, women are now bombarded with information and recommendations, which are often confusing at best and conflicting at worst. The objective of this review is to present current, evidence-based recommendations for some of the things that pregnant women should and should not routinely do during pregnancy.


Ultrasound equipment for the pediatric emergency department: a review.
Ultrasound is gaining momentum for use in the pediatric emergency department. It is important to understand the fundamentals of ultrasound equipment as it relates to pediatric emergency medicine.


MiRNA-708/CUL4B axis contributes into cell proliferation and apoptosis of osteosarcoma.
The functions of miRNA-708 for various diseases have been confirmed. However, its roles in osteosarcoma are unclear. In this study, we aimed to explore the role of miRNA-708 in osteosarcoma. Detection of the expression of miRNA-708 and CUL4B was used by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Cells were transfected with miRNA-708 mimics (mimics group) and miRNA negative control (NC group). Detection of cell growth curve at 24 h, 48 h, 72 h, and 96 h was made by cell counting kit-8 (CCK-8). Examination of the apoptosis rate was made by flow cytometry. The identification of the regulatory function was made by the luciferase reporter assay. The expression level of CUL4B was detected by Western blot. MiRNA-708 expression was reduced in the tumor cell lines. Compared with NC group, miRNA-708 expression was up-regulated by transfecting with mimics. Lower proliferation efficiency and higher cell apoptosis were showed in miRNA-708 mimics group relative to NC group. MiRNA-708 could regulate the expression of CUL4B by binding to its 3'UTR area. Furthermore, lower miRNA-708 and higher CUL4B were expressed in tumor tissues. MiRNA-708 expression was lower in tissues with IIB-III stage than that in IA-IIA stage. MiRNA-708/CUL4B axis contributes into cell proliferation and apoptosis of osteosarcoma.


Therapeutic Ultrasound in Navicular Stress Injuries in Elite Track and Field Athletes.
To ascertain whether therapeutic ultrasound (TUS) can be used to assess the progression of conservative management in navicular stress injuries. This is a prospective, clinical case series. Level of evidence IV. All participants were examined and followed up in a private Sports Injury Clinic. Ten elite track and field athletes with severe dorsal midfoot pain over the navicular bone participated in this study. All patients underwent both TUS and magnetic resonance imaging (MRI) evaluation. The painful threshold of TUS on initial evaluation was a mean of 0.707 ± 149 W/cm, and MRI detected a navicular stress injury in all patients. The athletes received conservative treatment and underwent sequential TUS evaluations at 4, 8, 12 and 16 weeks. Therapeutic ultrasound pain threshold values were recorded, and the patients were additionally asked to grade local tenderness on a Visual Analogue Scale. Time to return to play was also recorded. The level of pain produced by the application of TUS on a navicular stress fracture seemed to correlate well with Visual Analogue Scale scores and the grade of fracture demonstrated on MRI. The initial low TUS painful mean value increased to a normal mean value of 1.97 ± 0.067 W/cm by 16 weeks. When clinical and TUS findings had returned to normal, the patients were allowed to return to sports activities, with no recurrences experienced during the study period. The production of pain associated with the application of TUS on a navicular stress fracture is a safe and reproducible method of monitoring the resolution of these fractures. We have used it successfully in making return-to-play decisions for elite level track and field athletes.


Surgical management of the aging upper face.
Creating a more youthful appearance of the aging upper face requires a treatment plan which is tailored to address the patient's individual needs. The process begins with a comprehensive knowledge of anatomy and the physical effects of aging, involves discussion and understanding of aesthetic ideals, and culminates in the selection and performance of the appropriate procedure.


Initial invasion of gametophytic self-incompatibility alleles in the absence of tight linkage between pollen and pistil S alleles.
In homomorphic self-incompatibility (SI) systems of plants, the loci controlling the pollen and pistil types are tightly linked, and this prevents the generation of compatible combinations of alleles expressing pollen and pistil types, which would result in self-fertilization. We modeled the initial invasion of the first pollen and pistil alleles in gametophytic SI to determine whether these alleles can stably coexist in a population without tight linkage. We assume pollen and pistil loci each carry an incompatibility allele S and an allele without an incompatibility function N. We assume that pollen with an S allele are incompatible with pistils carrying S alleles, whereas other crosses are compatible. Ovules in pistils carrying an S allele suffer viability costs because recognition consumes resources. We found that the cost of carrying a pistil S allele allows pollen and pistil S alleles to coexist in a stable equilibrium if linkage is partial. This occurs because parents that carry pistil S alleles but are homozygous for pollen N alleles cannot avoid self-fertilization; however, they suffer viability costs. Hence, pollen N alleles are selected again. When pollen and pistil S alleles can coexist in a polymorphic equilibrium, selection will favor tighter linkage.


Distal embolization during lower extremity endovascular interventions.
Distal embolization (DE) during peripheral arterial endovascular interventions is a well-known complication that is poorly studied. The goal of this study was to determine the incidence, risk factors, and effect of DE on the outcomes of lower extremity endovascular interventions (LEIs). All LEIs between 2010 and 2014 in the Vascular Study Group of New England (VSGNE) database were reviewed. Patient characteristics were analyzed to determine predictors of DE. LEIs involving the superficial femoral artery (SFA) were reviewed to assess the effect of type of treatment on DE. The outcomes examined were loss of patency, limb loss, and mortality after LEI involving the SFA. A multivariable regression was used to determine predictors of DE. There were 10,875 procedures. The incidence of DE was 17.3 per 1000 procedures, and 68% required treatment (57% endovascular, 11% open surgery). DE was more common in patients treated for critical limb ischemia compared with claudication (relative risk [RR], 2.06; 95% confidence interval [CI], 1.24-3.45; P = .006) and for emergency interventions compared with elective (RR, 2.98; 95% CI, 1.22-7.30; P = .017). DE increased with the number of arteries treated (P < .0001) and with the length of occlusion (P < .0001). The SFA was the most commonly treated artery (4751 [43.7%]). In comparison with atherectomy and balloon angioplasty, stenting alone (RR, 0.36; 95% CI, 0.17-0.73; P = .005), balloon angioplasty alone (RR, 0.23; 95% CI, 0.13-0.41; P < .0001), and combined stenting and balloon angioplasty (RR, 0.29; 95% CI, 0.17-0.49; P < .0001) were associated with a significantly lower risk of DE. DE was not significantly associated with loss of patency, major amputation, or mortality. The incidence of DE during LEIs is 1% to 2% in the VSGNE database, and most patients are treated with additional endovascular interventions. The incidence increases in patients with critical limb ischemia and with the use of atherectomy.


Normalization with genes encoding ribosomal proteins but not GAPDH provides an accurate quantification of gene expressions in neuronal differentiation of PC12 cells.
Gene regulation at transcript level can provide a good indication of the complex signaling mechanisms underlying physiological and pathological processes. Transcriptomic methods such as microarray and quantitative real-time PCR require stable reference genes for accurate normalization of gene expression. Some but not all studies have shown that housekeeping genes (HGKs), beta-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which are routinely used for normalization, may vary significantly depending on the cell/tissue type and experimental conditions. It is currently unclear if these genes are stably expressed in cells undergoing drastic morphological changes during neuronal differentiation. Recent meta-analysis of microarray datasets showed that some but not all of the ribosomal protein genes are stably expressed. To test the hypothesis that some ribosomal protein genes can serve as reference genes for neuronal differentiation, a genome-wide analysis was performed and putative reference genes were identified based on stability of expressions. The stabilities of these potential reference genes were then analyzed by reverse transcription quantitative real-time PCR in six differentiation conditions. Twenty stably expressed genes, including thirteen ribosomal protein genes, were selected from microarray analysis of the gene expression profiles of GDNF and NGF induced differentiation of PC12 cells. The expression levels of these candidate genes as well as ACTB and GAPDH were further analyzed by reverse transcription quantitative real-time PCR in PC12 cells differentiated with a variety of stimuli including NGF, GDNF, Forskolin, KCl and ROCK inhibitor, Y27632. The performances of these candidate genes as stable reference genes were evaluated with two independent statistical approaches, geNorm and NormFinder. The ribosomal protein genes, RPL19 and RPL29, were identified as suitable reference genes during neuronal differentiation of PC12 cells, regardless of the type of differentiation conditions. The combination of these two novel reference genes, but not the commonly used HKG, GAPDH, allows robust and accurate normalization of differentially expressed genes during PC12 differentiation.


Increased serum endotoxin and elevated CD14 and IL-1beta expression in a rat model of cerebrogenic multiple organ dysfunction syndrome.
To study the mechanisms underlying cerebrogenic multiple organ dysfunction syndrome (CMODS) through investigation of endotoxin levels and the expression of endotoxin receptor CD14 and interleukin IL-1beta mRNAs in a rat CMODS model. Acute cerebral hemorrhage was induced in Wistar rats by focal intracerebral injection of collagenase into the caudate nucleus. Serum endotoxin levels were quantitated using a chromogenic limulus lysate method; CD14 endotoxin receptor mRNA and IL-1beta mRNA levels in lung and intestine were determined by in situ hybridization. Serum endotoxin levels increased after 12 h, reaching a peak after 24 h, and declined to control levels at 72 h. The increase was statistically significant (P<0.05) compared to unoperated controls and the sham-operated group respectively. CD14 mRNA in lung and intestine increased after 12 h, peaked after 24-36 h, and then declined after 48 h. IL-1beta mRNA levels were also increased in lung and intestine (P<0.05), peaking at 36 h and declining thereafter. Expression levels of both CD14 and IL-1beta mRNAs correlated significantly with serum endotoxin levels (Plt;0.01). We conclude that acute cerebral hemorrhage results in endotoxemia and widespread increases in CD14 and IL-1beta expression. We suggest that acute cerebrovascular challenge leads to a stress/shock response that compromises the intestinal mucosal barrier. In turn, this allows endotoxin translocation into the body that provokes the release of pro-inflammatory lymphokines, leading to a systemic inflammatory response syndrome (SIRS) that culminates in multiple organ dysfunction.


Synthesis, characterization and toxicity studies of pyridinecarboxaldehydes and L-tryptophan derived Schiff bases and corresponding copper (II) complexes.
Schiff bases and their metal-complexes are versatile compounds exhibiting a broad range of biological activities and thus actively used in the drug development process. The aim of the present study was the synthesis and characterization of new Schiff bases and their copper (II) complexes, derived from L-tryptophan and isomeric (2-; 3-; 4-) pyridinecarboxaldehydes, as well as the assessment of their toxicity in vitro. The optimal conditions of the Schiff base synthesis resulting in up to 75-85% yield of target products were identified. The structure-activity relationship analysis indicated that the location of the carboxaldehyde group at 2-, 3- or 4-position with regard to nitrogen of the pyridine ring in aldehyde component of the L-tryptophan derivative Schiff bases and corresponding copper complexes essentially change the biological activity of the compounds. The carboxaldehyde group at 2- and 4-positions leads to the higher cytotoxic activity, than that of at 3-position, and the presence of the copper in the complexes increases the cytotoxicity. Based on toxicity classification data, the compounds with non-toxic profile were identified, which can be used as new entities in the drug development process using Schiff base scaffold.


Delivering high-quality cardiopulmonary resuscitation in-hospital.
This review discusses recent data relating to delivering high-quality cardiopulmonary resuscitation (CPR) to patients with in-hospital cardiac arrest. Delivering high-quality CPR requires interventions at a national, local, team and individual rescuer level. These include measuring patient outcomes, patient safety incident reporting, education, an increased emphasis on human factors, briefing and debriefing of resuscitation teams, and the use of sensing devices that provide real-time prompts or feedback to rescuers during CPR. Data from national registries, patient safety incident reports and mock codes can be used to identify areas for improving practice. Education of staff is essential in both technical and nontechnical resuscitation skills (human factors). Resuscitation team performance can be improved by ensuring teams brief and plan beforehand and also debrief using feedback data collected during resuscitation events. The use of feedback and prompt devices helps improve adherence to guidelines for chest compression quality but data are lacking in terms of showing improved patient outcomes. Delivering high-quality CPR in-hospital requires a multifaceted approach. Collecting data during arrests and feeding back in real time and postevent during debriefings can be used to improve delivery of high-quality CPR. There are few studies that show improvement in actual patient outcomes (e.g., survival to hospital discharge) with improvements in delivery of high-quality CPR. Recognizing the importance of both technical and nontechnical skills (human factors) to deliver high-quality CPR is essential.


[Diffusion of water through the narrow pores of biomembranes].
The pulsed-gradient, nuclear magnetic resonance method has been used to study diffusion of water through the cell membranes. An analytical expression is derived for relative amplitude of nuclear spin-echo, describing the diffusion of water in membrane pores. The latter are so narrow, that their diameter is only a little greater than the water molecule diameter. The experimental data for consumed yeast with normal content of water agree well with the theoretical dependence.


Sequence-Specific Targeting of Bacterial Resistance Genes Increases Antibiotic Efficacy.
The lack of effective and well-tolerated therapies against antibiotic-resistant bacteria is a global public health problem leading to prolonged treatment and increased mortality. To improve the efficacy of existing antibiotic compounds, we introduce a new method for strategically inducing antibiotic hypersensitivity in pathogenic bacteria. Following the systematic verification that the AcrAB-TolC efflux system is one of the major determinants of the intrinsic antibiotic resistance levels in Escherichia coli, we have developed a short antisense oligomer designed to inhibit the expression of acrA and increase antibiotic susceptibility in E. coli. By employing this strategy, we can inhibit E. coli growth using 2- to 40-fold lower antibiotic doses, depending on the antibiotic compound utilized. The sensitizing effect of the antisense oligomer is highly specific to the targeted gene's sequence, which is conserved in several bacterial genera, and the oligomer does not have any detectable toxicity against human cells. Finally, we demonstrate that antisense oligomers improve the efficacy of antibiotic combinations, allowing the combined use of even antagonistic antibiotic pairs that are typically not favored due to their reduced activities.


Connexin distribution in the rabbit and rat ciliary body. A case for heterotypic epithelial gap junctions.
To evaluate the distribution of different alpha- and beta-type connexins (Cx) present in the dual layered ciliary body epithelia (CBE) of both rabbit and rat. Immunocytochemical detection of Cx26, Cx32, Cx43, and Cx50 was performed on frozen sections of rabbit and rat ciliary body using indirect immunofluorescent methods. The identity of the antigens recognized by the monoclonal primary antibodies was further confirmed by Western immunoblots. Double labeling experiments based on either conventional or confocal microscopy were carried out to establish the exact spatial relationship between different connexins. Connexin 50 was found only in the nonpigmented epithelium (NPE) at apical and basolateral membranes, whereas Cx43 was observed exclusively and at a very high concentration in the pigmented epithelium (PE), primarily in the apical cell membrane, with minimal extension to the proximal lateral zone. The correct antigenicity of the antibodies was confirmed by Western blots of rabbit ciliary body membranes. In rabbit, the Cx26 antibody detected an antigen that was abundant in the NPE and was weakly expressed in the PE. In rat, however, the Cx26 staining was confined to capillary wall endothelia. Western blots of ciliary body and liver membranes and liver immunohistology indicated that the Cx26 antibody used does not recognize rabbit Cx26. Cx32 did not yield any substantial epithelial labeling in either species. The distribution of Cx50 around the entire NPE cell perimeter suggests its involvement in NPE-NPE cell homotypic gap junctions. The concentration of Cx43 and Cx50 at the apical membranes of the PE and NPE cells, respectively, and their complete absence from the opposite cell suggest that these connexins may participate in the formation of heterotypic gap junctions, either with each other or with other yet unidentified connexins.


How do adolescents adjust to their parent's multiple sclerosis? An interview study.
The aim of this study was to explore how adolescents with a parent with multiple sclerosis (MS) adjust to their parents' illness. We used an inductive qualitative approach with open ended questions as this was appropriate to facilitate the development of a broad and child-centred understanding of how adolescents adjust and which resources they use to cope with the challenges that are associated with parental MS. Fifteen semi-structured face-to-face interviews were conducted. Interviews were audiotaped, transcribed, and analysed using inductive thematic analysis. Adolescents described both positive and negative experiences related to having a parent with MS. Benefits to having a parent with MS included reports of feeling more empathetic to others and more grown-up. Negative impacts included family tension, less time to spend with friends, and worries about the future. The support from the well parent, siblings, and friends were found to facilitate adolescents' adjustment. Adolescents assuming a parenting role and illness characteristics, such as illness deterioration, relapses and fatigue, challenged adolescents' adjustment to having a parent with MS. Some adolescents described adjusting well to having a parent with MS, while others appeared to have more difficulty. Whilst the severity of the parent's deterioration and symptoms appeared to play a role in adjustment, other potentially modifiable factors such as the lack of well parent's support, adolescents' increased parenting responsibilities, and family tension also posed barriers to adolescents' adjustment. Support interventions may be helpful for vulnerable adolescents, which consider both family and individual factors.


Condom use in penile-vaginal intercourse among Canadian adults: Results from the sex in Canada survey.
This paper examines condom use in penile-vaginal sexual intercourse among adults in Canada. The Sex in Canada survey is a national survey of Canadian adults, ages 18+ (N = 2,303). The online survey used quota-based population sample matching of 2016 census targets for gender, age, region, language, visible minority status, and education level. We report general patterns of self-reported condom use, as well as results from zero-inflated negative binomial regression models on the relationship between condom use and social location, relationship status, and sexual health. Condom use varies by gender, age, education, visible minority status, and relationship status. Use of condoms is related to the perception of risk of being diagnosed with a sexually transmitted infection in the next six months and to the experience of receiving lessons in condom use. No significant associations were found between condom use and region, rural/urban residence, income, or religion. Among men, but not women, condom use is associated with language preference, past diagnosis with a sexually transmitted infection, and self-reported sexual health. Canadian adults report using a condom in approximately 30% of their sexual encounters involving penile-vaginal sex. Condom use is highest among young adults. Single people use condoms more often than people with marital or common-law partners. Condom use is higher among those with higher levels of education, among people belonging to visible minorities relative to white people, and for men relative to women. People who think they are likely to be diagnosed with a sexually transmitted infection in the next six months are more likely to use condoms than those who do not.


Triple-vessel percutaneous coronary revascularization in situs inversus dextrocardia.
Dextrocardia with situs inversus occurs in approximately one in 10,000 individuals of whom 20% have primary ciliary dyskinesia inherited as an autosomal recessive trait. These patients have a high incidence of congenital cardiac disease but their risk of coronary artery disease is similar to that of the general population. We report what is, to our knowledge, the first case of total triple-vessel coronary revascularization by percutaneous stent implantation in a 79-year-old woman with situs inversus dextrocardia. We describe the successful use of standard diagnostic and interventional guide catheters with counter rotation and transversely inversed image acquisition techniques. The case also highlights that the right precordial pain may represent cardiac ischemia in this population.


Effects of cytokine antagonists on the hepatic acute-phase response.
Endotoxin-induced hepatic acute-phase protein synthesis has been thought to be primarily regulated through cytokines such as interleukin-1 (IL-1) and interleukin-6 (IL-6). Previously, it was found that a 23-kDa murine acute-phase protein, the lipopolysaccharide (LPS)-induced protein (LIP), was synthesized following treatment of hepatocytes in vitro with LPS. Since this protein was also induced by IL-1 and IL-6, the present studies were undertaken to determine if the effect of endotoxin was mediated through these cytokines. Primary cultures of murine hepatocytes were treated with LPS, IL-1, IL-6, or an LPS-stimulated macrophage supernatant in the presence or absence of the IL-1 receptor antagonist (IL-1 RA) and/or an anti-IL-6 antibody. The cells were then radiolabeled with [35S]methionine. LIP was detected by electrophoresis and autoradiography of the secreted proteins. In vitro, IL-1 RA completely inhibited the stimulation of LIP synthesis elicited by IL-1 and the macrophage supernatant, but did not affect LPS-stimulated synthesis of this protein. The anti-IL-6 antibody inhibited IL-6-triggered synthesis of LIP, but had no effect on LPS-stimulated synthesis. Hepatocytes isolated from mice treated in vivo with both IL-1 RA and LPS synthesized LIP to the same degree as hepatocytes isolated from mice treated with LPS alone. LPS-stimulated synthesis of LIP in vitro does not require IL-1 or IL-6 as an obligatory intermediate. These results are consistent with the hypothesis that endotoxin can directly stimulate hepatocyte acute-phase protein synthesis in the absence of cytokines.


Is it possible to automate the discovery of process maps for the time-driven activity-based costing method? A systematic review.
The main objective of this manuscript was to identify the methods used to create process maps for care pathways that utilized the time-driven activity-based costing method. This is a systematic mapping review. Searches were performed in the Embase, PubMed, CINAHL, Scopus, and Web of Science electronic literature databases from 2004 to September 25, 2022. The included studies reported practical cases from healthcare institutions in all medical fields as long as the time-driven activity-based costing method was employed. We used the time-driven activity-based costing method and analyzed the created process maps and a qualitative approach to identify the main fields. A total of 412 studies were retrieved, and 70 articles were included. Most of the articles are related to the fields of orthopedics and childbirth-related to hospital surgical procedures. We also identified various studies in the field of oncology and telemedicine services. The main methods for creating the process maps were direct observational practices, complemented by the involvement of multidisciplinary teams through surveys and interviews. Only 33% of the studies used hospital documents or healthcare data records to integrate with the process maps, and in 67% of the studies, the created maps were not validated by specialists. The application of process mining techniques effectively automates models generated through clinical pathways. They are applied to the time-driven activity-based costing method, making the process more agile and contributing to the visualization of high degrees of variations encountered in processes, thereby making it possible to enhance and achieve continual improvements in processes.


Alanine mutagenesis of high-mobility-group-protein-1 box B (HMG1-B).
We have generated a set of alanine-scanning substitutions in high-mobility-group protein 1 box B (HMG1-B; the second domain of the HMG1 nuclear protein from the rat) in order to explore the influence of specific surface side chains on its function and folding. Guanidine hydrochloride and thermal unfolding studies have been carried out to investigate the effect of substituted residues on the folding pathway. Binding to four-way junction and linear-duplex DNA has been assayed to determine which residues play an important role in DNA binding. We have identified several mutants that are more stable or bind more tightly to the junction than the wild-type, including the particular phenylalanine side chain that is thought to intercalate into the DNA. Thus the interaction between HMG1-B and branched DNA substrates should exhibit differences from present models based on the structure of the complexes that have been solved to date.


Therapeutic strategies against BK polyomavirus infection in kidney transplant recipients: Systematic review and meta-analysis.
The selection of antiviral therapy for BK polyomavirus (BKPyV) infection has been extensively debated. Our study aimed to assess the efficacy and safety of various treatments for BKPyV infection. We searched PubMed, EMBASE, and Web of Science databases for relevant studies regarding drug treatments for BKPyV viremia/DNAemia published between January 1, 1970 and September 30, 2022. Two independent authors screened the published studies, extracted pertinent data, and evaluated their methodological quality. A meta-analysis was performed using the RevMan software version 4.2.2. A total of 33 published studies involving 986 patients were included in the meta-analysis. Overall, therapeutic interventions comprised immunosuppression reduction alone or in combination with leflunomide, intravenous immunoglobulin (IVIG), cidofovir, or mTOR inhibitor (mTORi) therapy. The meta-analysis revealed that the efficacy of immunosuppression reduction alone for serum BKPyV clearance was 68% (95% confidence interval [CI]: 0.58-0.77; I2 = 78%). Moreover, the efficacy of immunosuppression reduction in combination with leflunomide, cidofovir, IVIG, or mTORi therapy for serum BKPyV clearance was 61% (95% CI: 0.47-0.74; I2 = 83%), 71% (95% CI: 0.63-0.78; I2 = 0), 87% (95% CI: 0.82-0.93; I2 = 45%), and 80% (95% CI: 0.59-1.00; I2 = 58%), respectively. Compared to immunosuppression reduction alone, immunosuppression reduction combined with IVIG therapy offered a statistically significant benefit in serum BKPyV clearance (P < 0.01) with minimal adverse reactions, whereas other adjunctive drug treatments did not demonstrate considerable effects. Reducing immunosuppression remains the primary approach for treating BKPyV infection. Although the combination treatment with IVIG proved to be most effective, other agents might offer varied antiviral advantages of high heterogeneity, which should be substantiated in future long-term randomized controlled trials.


The natural protective mechanism against hyperglycemia in vascular endothelial cells: roles of the lipid peroxidation product 4-hydroxydodecadienal and peroxisome proliferator-activated receptor delta.
Vascular endothelial cells (VECs) downregulate their rate of glucose uptake in response to hyperglycemia by decreasing the expression of their typical glucose transporter GLUT-1. Hitherto, we discovered critical roles for the protein calreticulin and the arachidonic acid-metabolizing enzyme 12-lipoxygenase in this autoregulatory process. The hypothesis that 4-hydroxydodeca-(2E,6Z)-dienal (4-HDDE), the peroxidation product of 12-lipoxygenase, mediates this downregulatory mechanism by activating peroxisome proliferator-activated receptor (PPAR) delta was investigated. Effects of 4-HDDE and PPARdelta on the glucose transport system and calreticulin expression in primary bovine aortic endothelial cells were evaluated by pharmacological and molecular interventions. Using GW501516 (PPARdelta agonist) and GSK0660 (PPARdelta antagonist), we discovered that high-glucose-induced downregulation of the glucose transport system in VECs is mediated by PPARdelta. A PPAR-sensitive luciferase reporter assay in VECs revealed that high glucose markedly increased luciferase activity, while GSK0660 abolished it. High-performance liquid chromatography analysis showed that high-glucose incubation substantially elevated the generation of 4-HDDE in VECs. Treatment of VECs, exposed to normal glucose, with 4-HDDE mimicked high glucose and downregulated the glucose transport system and increased calreticulin expression. Like high glucose, 4-HDDE significantly activated PPARdelta in cells overexpressing human PPAR (hPPAR)delta but not hPPARalpha, -gamma1, or -gamma2. Moreover, silencing of PPARdelta prevented high-glucose-dependent alterations in GLUT-1 and calreticulin expression. Finally, specific binding of PPARdelta to a PPAR response element in the promoter region of the calreticulin gene was identified by utilizing a specific chromatin immunoprecipitation assay. Collectively, our data show that 4-HDDE plays a central role in the downregulation of glucose uptake in VECs by activating PPARdelta.


Unintentional wildlife poisoning and proposals for sustainable management of rodents.
In Europe, bromadiolone, an anticoagulant rodenticide authorized for plant protection, may be applied intensively in fields to control rodents. The high level of poisoning of wildlife that follows such treatments over large areas has been frequently reported. In France, bromadiolone has been used to control water voles (Arvicola terrestris) since the 1980s. Both regulation and practices of rodent control have evolved during the last 15 years to restrict the quantity of poisoned bait used by farmers. This has led to a drastic reduction of the number of cases of poisoned wildlife reported by the French surveillance network SAGIR. During the autumn and winter 2011, favorable weather conditions and high vole densities led to the staging of several hundreds of Red Kites (Milvus milvus) in the Puy-de-Dôme department (central France). At the same time, intensive treatments with bromadiolone were performed in this area. Although no misuse has been mentioned by the authorities following controls, 28 Red Kites and 16 Common Buzzards (Buteo buteo) were found dead during surveys in November and December 2011. For all these birds, poisoning by bromadiolone as the main cause of death was either confirmed or highly suspected. Other observations suggest a possible impact of bromadiolone on the breeding population of Red Kites in this area during the spring 2011. French regulation of vole control for plant protection is currently under revision, and we believe this event calls for more sustainable management of rodent outbreaks. Based on large-scale experiments undertaken in eastern France, we propose that direct control of voles at low density (with trapping or limited chemical treatments) and mechanical destruction of vole tunnels, mole control, landscape management, and predator fostering be included in future regulation because such practices could help resolve conservation and agricultural issues.


Real world evidence for public health decision-making on vaccination policies: perspectives from an expert roundtable.
Influenza causes significant morbidity and mortality, but influenza vaccine uptake remains below most countries' targets. Vaccine policy recommendations vary, as do procedures for reviewing and appraising the evidence. During a series of roundtable discussions, we reviewed procedures and methodologies used by health ministries in four European countries to inform vaccine recommendations. We review the type of evidence currently recommended by each health ministry and the range of approaches toward considering randomized controlled trials (RCTs) and real-world evidence (RWE) studies when setting influenza vaccine recommendations. Influenza vaccine recommendations should be based on data from both RCTs and RWE studies of efficacy, effectiveness, and safety. Such data should be considered alongside health-economic, cost-effectiveness, and budgetary factors. Although RCT data are more robust and less prone to bias, well-designed RWE studies permit timely evaluation of vaccine benefits, effectiveness comparisons over multiple seasons in large populations, and detection of rare adverse events, under real-world conditions. Given the variability of vaccine effectiveness due to influenza virus mutations and increasing diversification of influenza vaccines, we argue that consideration of both RWE and RCT evidence is the best approach to more nuanced and timely updates of influenza vaccine recommendations.


[Results of autologous blood donation in orthopedic hip and knee joint replacement].
This study was undertaken to investigate the efficiency of autologous blood donation (ABD) with regard to saving of homologous transfusion, to determine the reasons for exclusion from donations and the rate of incidents during the procedure, and to investigate the quality of autologous fresh frozen plasma (AFFP). During an observation period of 4.5 years, all patients scheduled for elective orthopaedic hip and knee replacement were included. A period of 4 years was evaluated retrospectively, and the last half year was evaluated prospectively. Among a collective of 710 patients, 55 (8%) non-donors and 655 (92%) donors with a total number of 1592 ABD were found. Mean age of non-donors with hip surgery was significantly higher than that of donors (72 vs. 64 years), the same was observed in patients with knee surgery (71 vs. 68 years). In the hip surgery group, 11 of 338 patients were non-donors (3%), compared with 44 of 372 patients with knee surgery (12%). In the prospective part of the study, 7% of 121 patients were non-donors. Reasons for exclusion from donation were 5 times of medical and 4 times of organisational nature. In donors for hip surgery, a mean of 3.0 units was collected, compared with 1.9 units in donors for knee surgery. On the day before operation, mean haemoglobin concentrations were similar in donors and non-donors. During ABD, 11 incidents were observed, representing 0.69% of all ABD, 83.5% of 327 donors with hip surgery left the hospital without any transfusion of homologous blood, 16.5 of donors with hip surgery received one or more homologous transfusions, compared with 100% of non-donors (p < 0.001). In knee surgery, 93.3% of donors and 63.6% of non-donors required no homologous blood, whereas 6.7% of donors and 36.4% of non-donors received one or more homologous transfusions (p < 0.001). 529 of 2850 autologous blood units (19%) were not transfused, and 19 of these units were rejected due to technical or organisational problems. In 97 patients with 240 ABD and 240 AFFP, prothrombine time, fibrinogen concentration and AT III in defrosted AFFP exceeded 70% of the values determined before ABD. ABD is a safe procedure in almost all, even elderly, patients scheduled for elective orthopaedic hip or knee replacement in both types of operations, ABD reduces the risk of homologous transfusion significantly. A number of 3-4 units is necessary for total hip replacement, whereas 2 units are sufficient for partial or total knee replacement. Haemostatic quality of AFFP meets the requirements of fresh frozen homologous plasma.


Deconvolution of transcriptional networks identifies TCF4 as a master regulator in schizophrenia.
Applying tissue-specific deconvolution of transcriptional networks to identify their master regulators (MRs) in neuropsychiatric disorders has been largely unexplored. Here, using two schizophrenia (SCZ) case-control RNA-seq datasets, one on postmortem dorsolateral prefrontal cortex (DLPFC) and another on cultured olfactory neuroepithelium, we deconvolved the transcriptional networks and identified TCF4 as a top candidate MR that may be dysregulated in SCZ. We validated TCF4 as a MR through enrichment analysis of TCF4-binding sites in induced pluripotent stem cell (hiPSC)-derived neurons and in neuroblastoma cells. We further validated the predicted TCF4 targets by knocking down TCF4 in hiPSC-derived neural progenitor cells (NPCs) and glutamatergic neurons (Glut_Ns). The perturbed TCF4 gene network in NPCs was more enriched for pathways involved in neuronal activity and SCZ-associated risk genes, compared to Glut_Ns. Our results suggest that TCF4 may serve as a MR of a gene network dysregulated in SCZ at early stages of neurodevelopment.


Identification of GntR as regulator of the glucose metabolism in Pseudomonas aeruginosa.
In contrast to Escherichia coli, glucose metabolism in pseudomonads occurs exclusively through the Entner-Doudoroff (ED) pathway. This pathway, as well as the three routes to generate the initial ED pathway substrate, 6-phosphogluconate, is regulated by the PtxS, HexR and GtrS/GltR systems. With GntR (PA2320) we report here the identification of an additional regulator in Pseudomonas aeruginosa PAO1. GntR repressed its own expression as well as that of the GntP gluconate permease. In contrast to PtxS and GtrS/GltR, GntR did not modulate expression of the toxA gene encoding the exotoxin A virulence factor. GntR was found to bind to promoters PgntR and PgntP and the consensus sequence of its operator was defined as 5'-AC-N-AAG-N-TAGCGCT-3'. Both operator sites overlapped with the RNA polymerase binding site and we show that GntR employs an effector mediated de-repression mechanism. The release of promoter bound GntR is induced by gluconate and 6-phosphogluconate that bind with similar apparent affinities to the GntR/DNA complex. GntR and PtxS are paralogous and may have evolved from a common ancestor. The concerted action of four regulatory systems in the regulation of glucose metabolism in Pseudomonas can be considered as a model to understand complex regulatory circuits in bacteria.


Constitutive and inducible Saccharomyces cerevisiae promoters: evidence for two distinct molecular mechanisms.
his3 and pet56 are adjacent Saccharomyces cerevisiae genes that are transcribed in opposite directions from initiation sites that are separated by 200 base pairs. Under normal growth conditions, in which his3 and pet56 are transcribed at similar basal levels, a poly(dA-dT) sequence located between the genes serves as the upstream promoter element for both. In contrast, his3 but not pet56 transcription is induced during conditions of amino acid starvation, even though the critical regulatory site is located upstream of both respective TATA regions. Moreover, only one of the two normal his3 initiation sites is subject to induction. From genetic and biochemical evidence, I suggest that the his3-pet56 intergenic region contains constitutive and inducible promoters with different properties. In particular, two classes of TATA elements, constitutive (Tc) and regulatory (Tr), can be distinguished by their ability to respond to upstream regulatory elements, by their effects on the selection of initiation sites, and by their physical structure in nuclear chromatin. Constitutive and inducible his3 transcription is mediated by distinct promoters representing each class, whereas pet56 transcription is mediated by a constitutive promoter. Molecular mechanisms for these different kinds of S. cerevisiae promoters are proposed.


Field stitching approach for the wave optical modeling of black silicon structures.
The interest in black silicon structures as an anti-reflective interface at the front side of silicon solar cells increased strongly with the rise of diamond wire sawing. The application of optical modeling in order to predict optimal structure parameters could be highly valuable. However, due to the random nature of the structure as well as dimensions in the range of the wavelengths of interest, optical modeling is still a challenge. Within this work, the stitching method of rigorously calculated fields is extended and applied to a black silicon structure. A Fourier transform is used to determine the angular intensity distribution in the far field. In combination with the OPTOS formalism, this allows modeling of silicon substrates with black silicon front side and shows a reasonably good agreement with optical measurement results. Implementing the investigated structure into a solar cell configuration reveals not only a low reflectance but also a very good light trapping performance close to that of a Lambertian scatterer.


[Subcutaneous fibrosarcoma in a lion (Panthera leo)].
A subcutaneous growth developed on the left side of a zoo lion. The growth enlarged over a period of 3 months. A diagnosis of fibrosarcoma was made on account of the history, clinical and histological examinations. The lion was killed 3 weeks after surgical removal of the tumour. The autopsy showed no evidence of metastases. A foreign body (wire) was discovered in the abdominal cavity but no association was found between the lesion caused by the wire and the malignancy.


Risk and Benefit Profile of Dulaglutide in Established Therapeutic Indication.
Recently, Food and Drug Administration (FDA) has approved Dulaglutide (GLP-1 analog) for the treatment of type 2 diabetes mellitus. However, regarding Adverse Drug Reactions (ADRs) of Dulaglutide in a large group of population, very less information is available. Thus, in the present investigation, we tried to find out the risk and benefit profile of Dulaglutide. The data related to risk and benefit profile of Dulaglutide has been extracted from various sources like Pub-Med, Regulatory Websites of various agencies, Clinical trial registry etc. from Sep 2014 (first approval) to Feb 2018. A total of 182 studies have been published from Sep 2014 to Feb 2018 regarding Dulaglutide. After inclusion and exclusion criteria, 25 studies have been selected for risk and benefit analysis. The results of the current study have shown the various therapeutic benefits of Dulaglutide (reduces weight, decreases progression of nephropathy, reduces incidence of myocardial infarction, reduces blood pressure etc.) in the treatment of type 2 diabetes mellitus. However, emerging evidence has also indicated the various risks associated with Dulaglutide such as septicemia, malignant neoplasm, coronary artery disease and pancreatic cancer. In conclusion, Marketing Authorization Holder (MAH) and Regulatory Authorities (RAs) should consider these safety issues in future.


SRC Increases MYC mRNA Expression in Estrogen Receptor-Positive Breast Cancer via mRNA Stabilization and Inhibition of p53 Function.
The transcription factor gene MYC is important in breast cancer, and its mRNA is maintained at a high level even in the absence of gene amplification. The mechanism(s) underlying increased MYC mRNA expression is unknown. Here, we demonstrate that MYC mRNA was stabilized upon estrogen stimulation of estrogen receptor-positive breast cancer cells via SRC-dependent effects on a recently described RNA-binding protein, IMP1 with an N-terminal deletion (ΔN-IMP1). We also show that loss of the tumor suppressor p53 increased MYC mRNA levels even in the absence of estrogen stimulation. However, in cells with wild-type p53, SRC acted to overcome p53-mediated inhibition of estrogen-stimulated cell cycle entry and progression. SRC thus promotes cell proliferation in two ways: by stabilizing MYC mRNA and by inhibiting p53 function. Since estrogen receptor-positive breast cancers typically express wild-type p53, these studies establish a rationale for p53 status to be predictive for effective SRC inhibitor treatment in this subtype of breast cancer.


Development of multi-dimensional body image scale for malaysian female adolescents.
The present study was conducted to develop a Multi-dimensional Body Image Scale for Malaysian female adolescents. Data were collected among 328 female adolescents from a secondary school in Kuantan district, state of Pahang, Malaysia by using a self-administered questionnaire and anthropometric measurements. The self-administered questionnaire comprised multiple measures of body image, Eating Attitude Test (EAT-26; Garner & Garfinkel, 1979) and Rosenberg Self-esteem Inventory (Rosenberg, 1965). The 152 items from selected multiple measures of body image were examined through factor analysis and for internal consistency. Correlations between Multi-dimensional Body Image Scale and body mass index (BMI), risk of eating disorders and self-esteem were assessed for construct validity. A seven factor model of a 62-item Multi-dimensional Body Image Scale for Malaysian female adolescents with construct validity and good internal consistency was developed. The scale encompasses 1) preoccupation with thinness and dieting behavior, 2) appearance and body satisfaction, 3) body importance, 4) muscle increasing behavior, 5) extreme dieting behavior, 6) appearance importance, and 7) perception of size and shape dimensions. Besides, a multidimensional body image composite score was proposed to screen negative body image risk in female adolescents. The result found body image was correlated with BMI, risk of eating disorders and self-esteem in female adolescents. In short, the present study supports a multi-dimensional concept for body image and provides a new insight into its multi-dimensionality in Malaysian female adolescents with preliminary validity and reliability of the scale. The Multi-dimensional Body Image Scale can be used to identify female adolescents who are potentially at risk of developing body image disturbance through future intervention programs.


Correlation of glutamate plus aspartate dose, plasma amino acid concentration and neuronal necrosis in infant mice.
Eight-day-old mice were given by gavage glutamate and aspartate mixtures providing each amino acid at 125, 250 or 500 mg/kg body weight (250, 500 and 1000 mg total dicarboxylic amino acids/kg) and the degree and extent of neuronal necrosis were determined. Similar studies were carried out in mice given monosodium L-glutamate at 250 or 500 mg/kg body weight. Plasma aspartate and glutamate concentrations were determined at each dose level. No animal given either glutamate or the glutamate plus aspartate mixture at 250 mg/kg developed neuronal necrosis. However, neuronal necrosis developed in 30% of animals given glutamate at 500 mg/kg (12+/-2 necrotic neurons/section in the region of maximal damage) and in 17% of animals given 250 mg glutamate/kg plus 250 mg aspartate/kg (11-13 necrotic neurons/section in the region of maximal damage). The threshold mean peak plasma glutamate plus aspartate concentration associated with neuronal necrosis was 128+/-24 mumol/dl. Using these data, and previously published data for aspartate-induced neurotoxicity (Finkelstein et al. Toxicology 1983, 29, 109), the individual threshold plasma glutamate and aspartate concentrations associated with neuronal necrosis were calculated to be 110 mumol/dl for aspartate and 75 mumol/dl for glutamate.


Predictor variables associated with positive Fast Track outcomes at the end of third grade.
Progress has been made in understanding the outcome effects of preventive interventions and treatments designed to reduce children's conduct problems. However, limited research has explored the factors that may affect the degree to which an intervention is likely to benefit particular individuals. This study examines selected child, family, and community baseline characteristics that may predict proximal outcomes from the Fast Track intervention. The primary goal of this study was to examine predictors of outcomes after 3 years of intervention participation, at the end of 3rd grade. Three types of proximal outcomes were examined: parent-rated aggression, teacher-rated oppositional-aggressive behavior, and special education involvement. The relation between 11 risk factors and these 3 outcomes was examined, with separate regression analyses for the intervention and control groups. Moderate evidence of prediction of outcome effects was found, although none of the baseline variables were found to predict all 3 outcomes, and different patterns of prediction emerged for home versus school outcomes.


Management of acute retrobulbar haemorrhage: a survey of non-ophthalmic emergency department physicians.
Acute retrobulbar haemorrhage (RBH) with orbital compartment syndrome is a sight-threatening ophthalmic emergency requiring treatment with lateral canthotomy and cantholysis (LC/C). However, such cases may present to non-ophthalmic emergency departments (ED) out-of-hours, when specialist intervention is not readily available. We completed a survey of ED physicians to explore experiences of RBH and confidence in undertaking LC/C. From February to April 2018, an online survey was sent to ED physicians of all training grades in seven UK locations. The survey comprised a case vignette of a patient presenting with clinical features of RBH with orbital compartment syndrome, with multiple choice questions on the diagnosis, management and onward referral of such cases. Additional questions explored the experience of RBH, LC/C and perspectives on current and future training of ED physicians in this area. 190 ED doctors completed the survey (response rate 70%). While 82.8% correctly diagnosed RBH and 95.7% recognised irreversible visual loss as a consequence of untreated RBH with orbital compartment syndrome, 78.7% indicated that they would initially undertake CT imaging rather than performing LC/C. Only 38.9% had previously encountered a case of RBH and only 37.1% would perform LC/C themselves, with 91.4% indicating that this was due to lack of training. 92.2% felt that more training was required for ED physicians in RBH management and performing LC/C. While cases of RBH with orbital compartment syndrome are infrequent, it is important that RBH management with the vital, sight-saving skill of LC/C is added to the United Kingdom Royal College of Emergency Medicine training curriculum. At present, though the majority of ED physicians can identify RBH, the minority are willing or able to undertake LC/C, potentially risking irreversible but avoidable visual loss.


Abnormal expression of only the CD34 part of a transgenic CD34/herpes simplex virus-thymidine kinase fusion protein is associated with ganciclovir resistance.
Donor T cell alloreactivity can be efficiently controlled by retrovirus-mediated ex vivo transfer of a "suicide" gene encoding the wild-type herpes simplex virus thymidine kinase (wtHSV-tk) gene, allowing gene-modified cells (GMCs) to be sensitive to ganciclovir (GCV). A limitation to this approach was related to the presence of an inactive form of the wtHSV-tk gene, resulting from alternative splicing. A corrected HSV-tk (cHSV-tk) gene was developed in order to circumvent this problem and was fused to a truncated splice variant of the human CD34 molecule (tCD34) suitable for the selection of retrovirally transduced GMCs. We demonstrate now that, despite this correction, CD34-positive, but GCV-resistant, HUT and primary GMCs can still be generated after transduction with a retroviral vector encoding a tCD34/cHSV-tk fusion protein (FuProtein). Deletions in the HSV-tk part of the transgene account in part for this resistance. However, an additional mechanism involving proteolytic-dependent "breakage" of the FuProtein has been observed: the CD34 part of the FuProtein can be detected by Western blot, separated from its HSV-tk part. Although the HSV-tk protein alone is not detectable in GCV-resistant tCD34/cHSV-tk-transduced HUT cells, it can be detected in 293T cells transduced with another tCD34/HSVTK fusion vector, demonstrating that a posttranslational effect leads to the breakage of the FuProtein. This is to our knowledge the first example of a loss of function of a FuProtein, of which one part is still expressed while the other one, suffering a selection pressure, is no longer detectable.


Role of Tranexamic Acid in Reducing Blood Loss in Vaginal Delivery.
Anti-fibrinolytic agents are used to reduce obstetric blood loss as the fibrinolytic system is known to get activated after placental delivery. To evaluate the efficacy of parenteral tranexamic acid in reducing blood loss during normal labour and to compare it with the amount of blood loss in patients who received placebo in the third stage of labour. Patients with spontaneous labour or planned for induction of labour and fulfilling the inclusion criteria were recruited for the study. In each patient, the pre-delivery pulse rate, blood pressure, Hb gm% and PCV% were noted. Labour was monitored carefully using a partogram. The study group received Inj. Oxytocin and Inj. Tranexamic acid. The control group received Inj. Oxytocin and Placebo injection. Immediately after delivery of the baby, when all the liquor was drained, the patient was placed over a blood drape-a disposable conical, graduated plastic collection bag. The amount of blood collected in the blood drape was measured. Then the patient was given pre-weighed pads, which were weighed 2 h post-partum. The blood loss was measured by measuring the blood collected in the drape and by weighing the swabs before and after delivery. The total number of patients studied was 100-equally distributed in both the groups. The age group of the patients and BMI were comparable. There was a significant increase in the pulse rate and decrease in blood pressure in the control group as compared with the study group. The post-delivery haemoglobin and haematocrit were significantly reduced in the control group as compared to the study group. The mean blood loss at the end of 2 h was 105 ml in the study group and 252 ml in the control group. There was a significant increase in the usage of uterotonics and also in the need for blood transfusion in the control group; 12 % of the patients in the control group had to stay for more than 3 days compared to 2 % in the study group. Tranexamic acid injection, an antifibrinolytic agent when given prophylactically after the delivery of the baby, by intravenous route appears to reduce the blood loss and maternal morbidity during normal labour effectively.


Classification of aster yellows-group phytoplasmas based on combined analyses of rRNA and tuf gene sequences.
Seventy phytoplasma isolates, including 10 previously characterized reference strains, of the aster yellows group were examined by RFLP analysis of PCR-amplified rDNA and RFLP and sequence analysis of the tuf gene. On the basis of rDNA restriction profiles, seven previously proposed 16S rDNA subgroups (16SrI-A, -B, -C, -D, -E, -F and -K) were recognized in the material examined. In addition, three new subgroups that differ in the RFLP profiles were identified and designated 16SrI-L, 16SrI-M and 16SrI-N. Of the two types of rDNA sequences used, an 1800 bp fragment comprising the entire 16S rRNA gene and the 16S-23S rDNA spacer region proved more suitable for AY-group phytoplasma differentiation than a 1240 bp fragment of the 16S rRNA gene. Many differences in the rDNA profiles between the subgroups could be explained by sequence heterogeneity of the two phytoplasmal rRNA operons. The subgroups delineated by RFLP analysis of a 940 bp tuf gene fragment are consistent with subgroups defined on the basis of rDNA sequences. However, subgroups 16SrI-D, -L and -M showed the same tuf gene restriction profiles as subgroup 16SrI-B. This result was confirmed by sequence analysis in which these subgroups differed slightly in their tuf gene sequence, when compared with members of subgroup 16SrI-B. On the basis of combined analyses of rDNA and tuf gene sequences and in view of pathological aspects, the taxonomic distinction of AY-subgroups 16SrI-A, -B, -C, -D, -E, -F, -K and -N appears to be substantial.


Occupational Burnout among Otolaryngology-Head and Neck Surgery Trainees in Australia.
Surgical trainee burnout has gained attention recently as a significant factor leading to poorer quality of patient care, decreased productivity, and personal dysfunction. As a result, we aimed to determine the prevalence and associated risk factors for burnout among otolaryngology-head and neck surgery (OHNS) trainees in Australia. Cross-sectional survey. National cohort of accredited OHNS trainees in Australia. Participants completed the Maslach Burnout Inventory (MBI). Trainee burnout was defined if any threshold of the 3 MBI domains-emotional exhaustion, depersonalization, or personal accomplishment-reached an established high threshold. Demographic data on potential predictors of burnout, such as stressors, workload, satisfaction, and support systems, were collected from survey responses. Predictors were compared with the burnout status. Of 67 OHNS trainees, 60 responded (66.7% men). Burnout was common among respondents, with 73.3% suffering from burnout in at least 1 of the 3 MBI domains (70.0%, emotional exhaustion; 46.7%, depersonalization; 18.3%, personal accomplishment). Trainee burnout was significantly influenced by training location (chi-square, P = .05), living geographically apart from social supports (odds ratio [OR], 3.49; chi-square, P = .007), number of years trained rurally or away from social supports (Kendall's tau-B, P = .03), difficulty balancing work and nonwork commitments (OR, 10.0; chi-square, P = .03), training negatively affecting their partner or family (OR, 14.30; chi-square, P = .05), and feeling uncomfortable approaching a supervisor (OR, 2.50; chi-square, P < .0001). Burnout was found to be very common among OHNS trainees in Australia. The statistically significant predictors identified should be addressed to minimize trainee burnout.


Denoising Cough Sound Recordings Using Neural Networks.
Objective cough sound evaluation is useful in the diagnosis and management of respiratory diseases. However, the performance of cough sound analysis models can degrade in the presence of background noises common in everyday environments. This brings forward the need for cough sound denoising. This work utilizes a method for denoising cough sound recordings using signal processing and machine learning techniques, inspired by research in the field of speech enhancement. It uses supervised learning to find a mapping between the noisy and clean spectra of cough sound signals using a fully connected feed-forward neural network. The method is validated on a dataset of 300 manually annotated cough sound recordings corrupted with babble noise. The effect of various signal processing and neural network parameters on denoising performance is investigated. The method is shown to improve cough sound quality and intelligibility and outperform conventional denoising methods.


Effects of hypoxemia on the a- and b-waves of the electroretinogram in the cat retina.
Slow components of the electroretinogram (ERG) are sensitive to even mild hypoxemia (60 < P(a)O(2) < 100 mm Hg) in the cat eye. However, the electrical responses of the inner retina remain unchanged until P(a)O(2) is below 40 mm Hg. In this study, the effects of hypoxemia on photoreceptors, on which both slow ERG components and inner retinal activity depend, were examined by recording the a-wave of the ERG. The ERG of dark-adapted, anesthetized cats was recorded between an Ag-AgCl electrode in the vitreous humor and a reference electrode near the eye. Responses to bright flashes of diffuse white light were recorded at 3-minute intervals during hypoxemic episodes lasting 15 minutes to 2 hours. The cat a-wave was well described by the Lamb and Pugh a-wave model during normoxia and hypoxemia. During mild hypoxemia (P(a)O(2) of 50-60 mm Hg), small changes in a-wave amplitude were detected but did not become greater during severe hypoxemia. The mean decrease in the a-wave amplitude during severe hypoxemia (P(a)O(2) of 20-30 mm Hg) was 8.9% from the mean amplitude during air breathing. The effects of hypoxemia were more severe on the b-wave amplitude. The mean decrease in the b-wave was 35% at P(a)O(2) of 20-30 mm Hg. The a-wave is more resistant to severe hypoxemia than the b-wave. This implies that photoreceptor transduction works almost normally during hypoxemia and that failure of inner retinal PO(2) regulation causes the decrease in the b-wave. Previously observed changes in the amplitudes of slow ERG components during hypoxemia may result from changes in the ionic environment, rather than a failure of photoreceptor energy metabolism.


Moderate asthma: burden, mechanisms and therapeutic perspectives.
Global Initiative for Asthma (GINA) document provides a classification of asthma severity according with the current level of treatment required to achieve diseases control and underlines the limitations of this approach. In this review, we will provide an overview of recent investigations that have analyzed clinical and molecular features of moderate asthma. Moderate asthma is heterogeneous in terms of response to inhaled treatment and pathogenetic mechanisms underlying the clinical features. Analysis of inflammatory pathways in patients who do not achieve disease remission allows identification of patient subgroups that may benefit from specific biological treatments. Scientific progress makes increasingly clear that there are biological mechanisms capable of identifying and justifying the degree of severity of asthma. The identification of these, combined with the development of new pharmacological treatments, will be the cornerstones of improving the management of asthma in its degrees of severity.


Inhibition of mutant Kras and p53-driven pancreatic carcinogenesis by atorvastatin: Mainly via targeting of the farnesylated DNAJA1 in chaperoning mutant p53.
Recent studies have indicated that using statins to inhibit the mevalonate pathway induces mutant p53 degradation by impairing the interaction of mutant p53 with DnaJ subfamily A member 1 (DNAJA1). However, the role of the C-terminus of DNAJA1 with a CAAX box for farnesylation in the binding, folding, and translocation of client proteins such as mutant p53 is not known. In the present study, we used a genetically engineered mouse model of pancreatic carcinoma and showed that atorvastatin significantly increased animal survival and inhibited pancreatic carcinogenesis. There was a dramatic decrease in mutant p53 protein accumulation in the pancreatic acini, pancreas intraepithelial neoplasia lesions, and adenocarcinoma. Supplementation with farnesyl pyrophosphate, a substrate for protein farnesylation, rescued atorvastatin-induced mutant p53 degradation in pancreatic cancer cells. Tipifarnib, a farnesyltransferase inhibitor, mirrored atorvastatin's effects on mutant p53, degraded mutant p53 in a dose-dependent manner, and converted farnesylated DNAJA1 into unfarnesylated DNAJA1. Farnesyltransferase gene knockdown also significantly promoted mutant p53 degradation. Coimmunoprecipitation either by an anti-DNAJA1 or p53 antibody confirmed the direct interaction of mutant p53 and DNAJA1 and higher doses of atorvastatin treatments converted more farnesylated DNAJA1 into unfarnesylated DNAJA1 with much less mutant p53 pulled down by DNAJA1. Strikingly, C394S mutant DNAJA1, in which the cysteine of the CAAX box was mutated to serine, was no longer able to be farnesylated and lost the ability to maintain mutant p53 stabilization. Our results show that farnesylated DNAJA1 is a crucial chaperone in maintaining mutant p53 stabilization and targeting farnesylated DNAJA1 by atorvastatin will be critical for inhibiting p53 mutant cancer.


The periosteal requirement and temporal dynamics of BMP2-induced middle phalanx regeneration in the adult mouse.
Regeneration of mammalian limbs is restricted to amputation of the distal digit tip, the terminal phalanx (P3). The adjacent skeletal element, the middle phalanx (P2), has emerged as a model system to investigate regenerative failure and as a site to test approaches aimed at enhancing regeneration. We report that exogenous application of bone morphogenetic protein 2 (BMP2) stimulates the formation of a transient cartilaginous callus distal to the amputation plane that mediates the regeneration of the amputated P2 bone. BMP2 initiates a significant regeneration response during the periosteal-derived cartilaginous healing phase of P2 bone repair, yet fails to induce regeneration in the absence of periosteal tissue, or after boney callus formation. We provide evidence that a temporal component exists in the induced regeneration of P2 that we define as the "regeneration window." In this window, cells are transiently responsive to BMP2 after the amputation injury. Simple re-injury of the healed P2 stump acts to reinitiate endogenous bone repair, complete with periosteal chondrogenesis, thus reopening the "regeneration window" and thereby recreating a regeneration-permissive environment that is responsive to exogenous BMP2 treatment.


Assessing Change Over Time in Voice Handicap and Voice-Related Perceived Control Using Ecological Momentary Assessment.
Voice handicap has generally been measured at a single timepoint. Little is known about its variability from hour to hour or day to day. Voice handicap has been shown to be negatively related to voice-related perceived control in cross-sectional studies, but the within-person variability in voice-related perceived control is also unknown. We aimed to use ecological momentary assessment (EMA) to (1) assess the feasibility of EMA to examine daily voice handicap and voice-related perceived control in patients with voice disorders, (2) measure within-person variability in daily voice handicap and perceived control, and (3) characterize temporal associations (eg, correlations over time) between daily voice handicap and perceived control. Adults with voice problems were recruited from a large public university medical center in the Midwest. They completed baseline measures, followed by twice-daily assessments, including selected items measuring voice handicap and perceived control, and then repeated the baseline measures at the final timepoint. Feasibility was assessed via completion rates. Within-person variability was measured using standard deviations. Temporal associations were characterized using simulation modeling analysis. EMA of voice handicap and perceived control was feasible in this patient population. Momentary voice handicap varied more than perceived control, though both were variable. Multiple patterns of temporal associations between daily voice handicap and perceived control were found. These findings identified important variability in (1) measures of voice handicap and perceived control and (2) their associations over time. Future EMA studies in patients with voice disorders are both feasible and warranted.


Progesterone receptor membrane component 1 facilitates Ca2+ signal amplification between endosomes and the endoplasmic reticulum.
Membrane contact sites (MCSs) between endosomes and the endoplasmic reticulum (ER) are thought to act as specialized trigger zones for Ca2+ signaling, where local Ca2+ released via endolysosomal ion channels is amplified by ER Ca2+-sensitive Ca2+ channels into global Ca2+ signals. Such amplification is integral to the action of the second messenger, nicotinic acid adenine dinucleotide phosphate (NAADP). However, functional regulators of inter-organellar Ca2+ crosstalk between endosomes and the ER remain poorly defined. Here, we identify progesterone receptor membrane component 1 (PGRMC1), an ER transmembrane protein that undergoes a unique heme-dependent dimerization, as an interactor of the endosomal two pore channel, TPC1. NAADP-dependent Ca2+ signals were potentiated by PGRMC1 overexpression through enhanced functional coupling between endosomal and ER Ca2+ stores and inhibited upon PGRMC1 knockdown. Point mutants in PGMRC1 or pharmacological manipulations that reduced its interaction with TPC1 were without effect. PGRMC1 therefore serves as a TPC1 interactor that regulates ER-endosomal coupling with functional implications for cellular Ca2+ dynamics and potentially the distribution of heme.


An efficient density-functional-theory force evaluation for large molecular systems.
An efficient, linear-scaling implementation of Kohn-Sham density-functional theory for the calculation of molecular forces for systems containing hundreds of atoms is presented. The density-fitted Coulomb force contribution is calculated in linear time by combining atomic integral screening with the continuous fast multipole method. For higher efficiency and greater simplicity, the near-field Coulomb force contribution is calculated by expanding the solid-harmonic Gaussian basis functions in Hermite rather than Cartesian Gaussians. The efficiency and linear complexity of the molecular-force evaluation is demonstrated by sample calculations and applied to the geometry optimization of a few selected large systems.


High frequency of germ-line BRCA2 mutations among Hungarian male breast cancer patients without family history.
To determine the contribution of BRCA1 and BRCA2 mutations to the pathogenesis of male breast cancer in Hungary, the country with the highest male breast cancer mortality rates in continental Europe, a series of 18 male breast cancer patients and three patients with gynecomastia was analyzed for germ-line mutations in both BRCA1 and BRCA2. Although no germ-line BRCA1 mutation was observed, 6 of the 18 male breast cancer cases (33%) carried truncating mutations in the BRCA2 gene. Unexpectedly, none of them reported a family history for breast/ovarian cancer. Four of six truncating mutations were novel, and two mutations were recurrent. Four patients (22%) had a family history of breast/ovarian cancer in at least one first- or second-degree relative; however, no BRCA2 mutation was identified among them. No mutation was identified in either of the genes in the gynecomastias. These results provide evidence for a strong genetic component of male breast cancer in Hungary.


3D printed optical concentrators for LED arrays.
Additive manufacturing methods based on photopolymerization offer a promising potential for fabrication of high quality, highly transparent optical components. One use of these technologies involves fabrication of parts for very specific and narrow applications. In this work, we first performed optical raytracing simulations to model an optimized freeform nonimaging concentrator for a custom-built 12-LED array and then fabricated several waveguide concentrators using 3D printing and characterized their optical characteristics. Our results demonstrate that realizing an irradiance of 17 kW/m2 or more with an irradiance nonuniformity of better than 2 % over an area approaching 1 cm2 is realistic and that such an approach can rival intensities achieved with powerful lasers over a similar area. We also discuss an application where eight different types of LEDs were coupled into the waveguides to construct a solar simulator.


A predictive model for patients with median arcuate ligament syndrome.
Due to the rarity of median arcuate ligament (MAL) syndrome, patient selection for surgery remains difficult. This study provides a predictive model to optimize patient selection and predict outcomes following a MAL release. Prospective data from patients undergoing a MAL release included demographics, radiologic studies, and SF-36 questionnaires. Successful postoperative changes in SF-36 was defined as an improvement > 10% in the total SF-36 score. A logistic regression model was used to develop a clinically applicable table to predict surgical outcomes. Celiac artery (CA) blood flow velocities were compared pre- and postoperatively and Pearson correlations were examined between velocities and SF-36 score changes. 42 patients underwent a laparoscopic MAL release with a mean follow-up of 28.5 ± 18.8 months. Postoperatively, all eight SF-36 scales improved significantly. The logistic regression model for predicting surgical benefit was significant (p = 0.0244) with a strong association between predictors and outcome (R2 = 0.36). Age and baseline CA expiratory velocity were significant predictors of improvement and predicted clinical improvement. There were significant differences between pre- and postoperative CA velocities. Postoperatively, the bodily pain scale showed the most significant increase (64%, p < 0.0001). A table was developed using age and preoperative CA expiratory velocities to predict clinical outcomes. Laparoscopic MAL produces significant symptom improvement, particularly in bodily pain. This is one of the first studies that uses preoperative data to predict symptom improvement following a MAL release. Age and baseline CA expiratory velocity can be used to guide postoperative expectations in patients with MAL syndrome.


Identification and Validation of a Prognostic lncRNA Signature for Hepatocellular Carcinoma.
Background: An accumulating body of evidence suggests that long non-coding RNAs (lncRNAs) can serve as potential cancer prognostic factors. However, the utility of lncRNA combinations in estimating overall survival (OS) for hepatocellular carcinoma (HCC) remains to be elucidated. This study aimed to construct a powerful lncRNA signature related to the OS for HCC to enhance prognostic accuracy. Methods: The expression patterns of lncRNAs and related clinical data of 371 HCC patients were obtained based on The Cancer Genome Atlas (TCGA). Differentially expressed lncRNAs (DElncRNAs) were acquired by comparing tumors with adjacent normal samples. lncRNAs displaying significant association with OS were screened through univariate Cox regression analysis and the least absolute shrinkage and selection operator (LASSO) algorithm. All cases were classified into the validation or training group at the ratio of 3:7 to validate the constructed lncRNA signature. Data from the Gene Expression Omnibus (GEO) were used for external validation. We conducted real-time polymerase chain reaction (PCR) and assays for Transwell invasion, migration, CCK-8, and colony formation to determine the biological roles of lncRNA. Gene set enrichment analysis (GSEA) of the lncRNA model risk score was also conducted. Results: We identified 1292 DElncRNAs, among which 172 were significant in univariate Cox regression analysis. In the training group (n = 263), LASSO regression analysis confirmed 11 DElncRNAs including AC010547.1, AC010280.2, AC015712.7, GACAT3 (gastric cancer associated transcript 3), AC079466.1, AC089983.1, AC051618.1, AL121721.1, LINC01747, LINC01517, and AC008750.3. The prognostic risk score was calculated, and the constructed risk model showed significant correlation with HCC OS (log-rank P-value of 8.489e-9, hazard ratio of 3.648, 95% confidence interval: 2.238-5.945). The area under the curve (AUC) for this lncRNA model was up to 0.846. This risk model was confirmed in the validation group (n = 108), the entire cohort, and the external GEO dataset (n = 203). GACAT3 was highly expressed in HCC tissues and cell lines. Based on online databases, GACAT3 expression independently affects both OS and disease-free survival in HCC patients. Silencing GACAT3 in vitro significantly suppressed HCC cell proliferation, invasion, and migration. Moreover, pathways related to the lncRNA model risk score were confirmed by GSEA. Conclusion: The lncRNA signature established in this study can be used to predict HCC prognosis, which could provide novel clinical evidence to guide targeted HCC treatment.