Placental proteome in late‑onset of fetal growth restriction.
Fetal growth restriction (FGR) occurs when the fetus does not reach its genetically programmed intrauterine potential for growth and affects ~5‑10% of pregnancies. This condition is one of the leading causes of perinatal mortality and morbidity associated with obstetric and neonatal complications. Placental dysfunction in FGR causes an impairment in the transfer of nutrients and oxygen from the mother to the developing fetus. Maternal adaptations to placental insufficiency may also play a role in the pathophysiology of FGR. The present study aimed to compare the proteome of the placentas of 18 women with the physiological course of pregnancy and eutrophic fetus [estimated fetal weight (EFW) >10th percentile; control group] and 18 women with late FGR (EFW <10th percentile) diagnosed after 32 weeks of pregnancy, according to the Delphi consensus (study group). The U. Mann‑Whitney test was used to compare two independent groups. The R. Spearman correlation coefficient significance test was used to assess the existence of a relationship between the analyzed measurable parameters. P<0.05 was considered to indicate a statistically significant difference. The tests showed the presence of 356 different proteins which were responsible for the regulation of gene transcription control, inhibiting the activity of proteolytic enzymes, regulation of trophoblast proliferation and angiogenesis and inflammatory response. In the FGR placental proteome, other detected proteins were mostly involved in response to oxidative stress, cellular oxidation and detoxication, apoptosis, hemostatic and catabolic processes, energy transduction protein interactions, cell proliferation, differentiation and intracellular signaling. The present study used chromatographic mass‑spectrometry to compare the placental proteome profiles in pregnancies complicated by late‑onset FGR and normal pregnancy. Comparative analysis of proteomes from normal and FGR placentas showed significant differences. Further research is needed to clarify maternal and fetal adaptations to FGR.


Narratives of hospital health care professionals during the pandemic in the Northern Philippines: A phenomenological study.
Hospital health care professionals (HCPs) play a vital and crucial role in saving the lives of patients afflicted with the coronavirus disease 2019 (COVID-19). As the incidence of the disease keeps increasing, health care workers in hospitals face difficulties in discharging their duties. This study aimed to describe the lived experiences of HCPs. Particularly, it determined their challenges, motivations and coping strategies to craft grassroots policies for the enhancement of health care delivery in the Northern Philippines. Employing a phenomenological study design, 24 study participants were purposively selected and their narratives were elicited through focus group discussion. Responses were transcribed verbatim. After data analyses, the challenges, coping strategies and motivations of HCPs were identified. Results revealed that HCPs experienced a lot of challenges. Some of these were brought by the rapid changes in their environment as HCPs. Moreover, there was lack of comprehensive strategies that made them unprepared along with a lack of human and material resources. Furthermore, they experienced physical fatigue because of overwhelming workload, anxiety, fear and discrimination that led to depression. Despite these difficulties, they remain resilient because of religious coping, being true to their duty as HCPs and the support they receive from their fellows. Thus, the study recommends that protocols to ensue should centre on adequate pandemic preparedness and capacitation of HCPs.


Changes in energy metabolism of Mycobacterium tuberculosis in mouse lung and under in vitro conditions affecting aerobic respiration.
Transcription profiling of genes encoding components of the respiratory chain and the ATP synthesizing apparatus of Mycobacterium tuberculosis was conducted in vivo in the infected mouse lung, and in vitro in bacterial cultures subjected to gradual oxygen depletion and to nitric oxide treatment. Transcript levels changed dramatically as infection progressed from bacterial exponential multiplication (acute infection) to cessation of bacterial growth (chronic infection) in response to host immunity. The proton-pumping type-I NADH dehydrogenase and the aa3-type cytochrome c oxidase were strongly down-regulated. Concurrently, the less energy-efficient cytochrome bd oxidase was transiently up-regulated. The nitrate transporter NarK2 was also up-regulated, indicative of increased nitrate respiration. The reduced efficiency of the respiratory chain was accompanied by decreased expression of ATP synthesis genes. Thus, adaptation of M. tuberculosis to host immunity involves three successive respiratory states leading to decreased energy production. Decreased bacterial counts in mice infected with a cydC mutant (defective in the cytochrome bd oxidase-associated transporter) at the transition to chronic infection provided initial evidence that the bd oxidase pathway is required for M. tuberculosis adaptation to host immunity. In vitro, NO treatment and hypoxia caused a switch from transcription of type I to type II NADH dehydrogenase. Moreover, cytochrome bd oxidase expression increased, but cytochrome c oxidase expression decreased slightly (nitric oxide) or not at all (hypoxia). These specific differences in respiratory metabolism during M. tuberculosis growth arrest in vitro and in vivo will guide manipulation of in vitro conditions to model bacterial adaptation to host immunity.


Monitoring vaginal epithelial thickness changes noninvasively in sheep using optical coherence tomography.
High-resolution optical coherence tomography can be used noninvasively to evaluate vaginal morphologic features, including epithelial thickness, to assess this protective barrier in transmission of sexually transmitted infections and to monitor tissue response to topical medications and hormonal fluctuations. We examined the use of optical coherence tomography to measure epithelial thickness noninvasively before and after topical treatment with a drug that causes epithelial thinning. Twelve female sheep were treated with intravaginal placebo (n = 4) or nonoxynol-9 (n = 8). Vaginal optical coherence tomography images were obtained before and 24 hours after treatment. Four sheep in the nonoxynol-9 group were also examined on days 3 and 7. Vaginal biopsies were obtained on the last examination day. Epithelial thickness was measured in optical coherence tomography images and in hematoxylin and eosin-stained histologic sections from biopsies. Statistical analysis was performed using analyses of variance (significance P < .05). Baseline optical coherence tomography epithelial thickness measurements were similar (85 ± 19 μm placebo, 78 ± 20 μm nonoxynol-9; P = .52). Epithelial thinning was significant after nonoxynol-9 (32 ± 22 μm) compared with placebo (80 ± 15 μm) 24 hours after treatment (P < .0001). In the 4 nonoxynol-9-treated sheep followed for 7 days, epithelial thickness returned to baseline by day 3, and increased significantly on day 7. Epithelial thickness measurements from histology were not significantly different than optical coherence tomography (P = .98 nonoxynol-9, P = .93 hydroxyethyl cellulose). Drug-induced changes in the epithelium were clearly detectable using optical coherence tomography imaging. Optical coherence tomography and histology epithelial thickness measurements were similar, validating optical coherence tomography as a noninvasive method for epithelial thickness measurement, providing an important tool for quantitative and longitudinal monitoring of vaginal epithelial changes.


Research Perspectives on the Regulation and Physiological Functions of FGF21 and its Association with NAFLD.
Fibroblast growth factor 21 (FGF21) is a metabolic hormone primarily secreted from the liver and functions in multiple tissues. Various transcription factors induce FGF21 expression in the liver, which indicates that FGF21 is a mediator of multiple environmental cues. FGF21 alters metabolism under starvation conditions, protects the body from energy depletion, and extends life span. Pharmacological administration of FGF21 alleviates dyslipidemia and induces weight loss in obese animals. In addition to the well-studied functions of FG21, several lines of recent evidence indicate a possible link between FGF21 and non-alcoholic fatty liver disease (NAFLD). High serum levels of FGF21 are associated with NAFLD and its risk factors, such as endoplasmic reticulum stress and chronic inflammation. In addition, FGF21 alleviates the major risk factors of NAFLD, including obesity, dyslipidemia, and insulin insensitivity. Thus, FGF21 is a potential drug candidate for diseases, such as NAFLD, dyslipidemia, and type 2 diabetes. In this review, the research perspectives of FGF21 and therapeutic potencies of FGF21 as a modulator of NAFLD are summarized.


Photochemical formation and reversible base-induced cleavage of a phosphagallene.
The reactivity of Cp*Ga (Cp* = C5Me5) towards phosphanylidenephosphoranes of the type ArTerP(PMe3) (ArTer = DipTer 2,6-(2,6-iPr2C6H3)2C6H3), TipTer 2,6-(2,4,6-iPr3C6H2)2C6H3 was investigated. While no thermal reaction was observed (in line with DFT results), irradiation at 405 nm at low temperatures resulted in the formation of phosphagallenes DipTerP = GaCp* (1a) and TipTerP = GaCp* (1b) accompanied by release of PMe3. When warming the reaction mixture to ambient temperatures without irradiation, the clean re-formation of ArTerP(PMe3) and Cp*Ga in a second-order reaction was observed. Upon removal of PMe3, 1a and 1b were isolated and fully characterized. Both derivatives were found to be labile and decomposed to the phosphafluorenes 2a and 2b, indicating generation of the transient phosphinidene ArTerP along with Cp*Ga. First reactivity studies show that CO2 and H2O cleanly reacted with 1a, affording DipTerPCO (3) and DipTerPH2 (4), respectively.


A zinc finger protein, interacted with cyclophilin, affects root development via IAA pathway in rice.
The plant hormone auxin plays a crucial role in lateral root development. To better understand the molecular mechanisms underlying lateral root formation, an auxin-responsive gene OsCYP2 (Os02g0121300) was characterized from rice. Compared to the wild type, OsCYP2-RNAi (RNA interference) lines exhibited distinctive defects in lateral root development. Yeast two-hybrid and glutathione S-transferase pull-down results confirmed that OsCYP2 interacted with a C2HC-type zinc finger protein (OsZFP, Os01g0252900) which is located in the rice nucleus. T2 OsZFP-RNAi lines had significantly fewer lateral roots than did wild-type plants, which suggests a role for OsCYP2 and OsZFP in regulating lateral root development. Quantitative real-time polymerase chain reaction showed that the expression of certain Aux/IAA (auxin/indole-3-acetic acid) genes was altered in OsCYP2- and OsZFP-RNAi lines in response to IAA. These findings imply that OsCYP2 and OsZFP participate in IAA signal pathways controlling lateral root development. More importantly, OsIAA11 showed functional redundancy not only in OsCYP2-RNAi lines but also in OsZFP-RNAi lines, which provides important clues for the elucidation of mechanisms controlling lateral root development in response to auxin.


Exploring factors influencing alcohol consumption patterns among Chinese and Caucasians.
Past research on alcohol consumption patterns comparing Chinese and Caucasians indicates that significant group differences exist. Chinese, as a group, consume significantly less alcohol than their Caucasian counterparts. Explanations for these differences have been controversial. Some argue that it is due to cultural differences, whereas others contend that it is because of physiological differences. The present study explores this controversy using a new approach: model testing via path analysis. With a sample size of 178 Canadian Chinese and 161 Caucasian university students, the present study found that in both ethnic groups, cultural norms rather than physical symptoms were a significant predictor of alcohol consumption patterns. This finding strongly suggests that alcohol consumption behavior can be better understood by a social/psychological rather than a biomedical approach. To effect changes in alcohol assumption patterns, it is suggested that cultural interpretations of alcohol use should be examined.


Photocatalytic reduction of carbon dioxide by hydrous hydrazine over Au-Cu alloy nanoparticles supported on SrTiO3/TiO2 coaxial nanotube arrays.
Efficient photocatalytic conversion of CO2 into CO and hydrocarbons by hydrous hydrazine (N2H4⋅H2O) is achieved on SrTiO3/TiO2 coaxial nanotube arrays loaded with Au-Cu bimetallic alloy nanoparticles. The synergetic catalytic effect by the Au-Cu alloy nanoparticles and the fast electron-transfer in SrTiO3/TiO2 coaxial nanoarchitecture are the main reasons for the efficiency, while N2H4⋅H2O as the H source and electron donor provides a reducing atmosphere to protect the surface Cu atoms from oxidation, therefore maintaining the alloying effect which is the basis for the high photocatalytic activity and stability. This approach opens a feasible route to enhance the photocatalytic efficiency, which also benefits the development of photocatalysts and co-catalysts.


Re-operation for recurrent coronary artery and graft disease. A review of 73 patients in a group of 2573 consecutive first operations.
Between January 1980 and December 1986, 2573 patients underwent simple first time coronary artery bypass grafting, of whom 73 (65 males and 8 females) aged 34-69 years (mean 51.3 yrs) had repeat bypass grafts at Wythenshawe Hospital, Manchester. Of these 73 patients, 15 had a previous myocardial infarction, 5 hyperlipidaemia, 4 systemic hypertension, and 12 had a strong family history of ischemic heart disease. There was an overall deterioration of left ventricular function at the time of reoperation. The interval between the two operations was 5-131 months (mean 34.2 mths); recurrence of angina occurred earlier (mean 18.4 mths). Vessels grafted at the first operation were LAD (59), RCA (46), circumflex (41) and diagonal (13). The corresponding data at reoperation were LAD (55), RCA (46), circumflex (28) and diagonal (10). Blocked grafts were seen in 67 patients and new lesions noticed in 29. Reoperation was done using saphenous vein (129), internal mammary artery (5), arm veins (2) and tubular Gortex grafts (2). One patient had concurrent excision of a left ventricular aneurysm. Coronary anastomoses were performed with elective ventricular fibrillation (47) or cardioplegic arrest (91). Aortic cross clamp time varied from 0-92 minutes. Seven patients required intra-aortic balloon support. These patients died in the first 30 days, an operative mortality rate of 4.1%, and two 18 months after surgery. Sixty-eight percent of patients seen at 1 year were totally symptom free. We conclude that reoperation for coronary artery disease can be done with a low mortality and good immediate relief of symptoms.(ABSTRACT TRUNCATED AT 250 WORDS)


An age-structured model of hiv infection that allows for variations in the production rate of viral particles and the death rate of productively infected cells.
Mathematical models of HIV-1 infection can help interpret drug treatment experiments and improve our understanding of the interplay between HIV-1 and the immune system. We develop and analyze an age- structured model of HIV-1 infection that allows for variations in the death rate of productively infected T cells and the production rate of viral particles as a function of the length of time a T cell has been infected. We show that this model is a generalization of the standard differential equation and of delay models previously used to describe HIV-1 infection, and provides a means for exploring fundamental issues of viral production and death. We show that the model has uninfected and infected steady states, linked by a transcritical bifurcation. We perform a local stability analysis of the nontrivial equilibrium solution and provide a general stability condition for models with age structure. We then use numerical methods to study solutions of our model focusing on the analysis of primary HIV infection. We show that the time to reach peak viral levels in the blood depends not only on initial conditions but also on the way in which viral production ramps up. If viral production ramps up slowly, we find that the time to peak viral load is delayed compared to results obtained using the standard (constant viral production) model of HIV infection. We find that data on viral load changing over time is insufficient to identify the functions specifying the dependence of the viral production rate or infected cell death rate on infected cell age. These functions must be determined through new quantitative experiments.


A test of alternative explanations for the stage-like progression of adolescent substance use in four national samples.
Previous research has established that there is a stage-like phenomenon of adolescent substance use such that alcohol use usually precedes marijuana use, and marijuana use usually precedes hard drug use. The current study tests three potential explanations for this stage-like phenomenon using nonstandard log-linear models. In four national surveys dating from 1974 to 1988, the results were consistent for both sexes, ages ranging from 12 through 18, and for Whites, Blacks, and Hispanics. The results suggest that adolescents are more likely to use alcohol before marijuana, and marijuana before hard drugs because these substances are more widely used and are used at early ages. An additional reason for the stage-like phenomenon is that there is also a group of adolescents who are willing to try all types of substances and a second group who are unwilling to use any substances. Implications for future longitudinal studies are discussed.


Gene regulation in plasmid RK2: positive control by korA in the expression of korC.
The broad-host-range plasmid RK2 encodes three host-lethal kil genes whose actions are controlled by specific kor genes. We have shown previously that the 0' to 5.5' region of RK2 encodes both kilA and korC. Because of the lethal effect of kilA, plasmids with this region cannot be maintained in Escherichia coli unless the RK2 korA gene is also present. To investigate korC in the absence of kilA and therefore of korA, we first mapped kilA and korC to specific segments of the cloned 0' to 5.5' region. This allowed us to construct a korC+ plasmid missing the kilA region and thereby removed the need to have korA in the cell. We found that this korC-encoding plasmid alone is insufficient to control kilC. The korA function is required, and it can be supplied in trans. We also constructed a kilA+ korC- plasmid and found that korA is sufficient to control kilA. Thus, in addition to acting negatively to control kilA, korA acts positively to allow korC control of kilC. This korA dependence of korC is bypassed in a rho-115 mutant of E. coli. We consider the possibility that korA product acts as an antiterminator of transcription in korC expression.


A cortical locus for anisotropic overlay suppression of stimuli presented at fixation.
Human contrast sensitivity for narrowband Gabor targets is suppressed when superimposed on narrowband masks of the same spatial frequency and orientation (referred to as overlay suppression), with suppression being broadly tuned to orientation and spatial frequency. Numerous behavioral and neurophysiological experiments have suggested that overlay suppression originates from the initial lateral geniculate nucleus (LGN) inputs to V1, which is consistent with the broad tuning typically reported for overlay suppression. However, recent reports have shown narrowly tuned anisotropic overlay suppression when narrowband targets are masked by broadband noise. Consequently, researchers have argued for an additional form of overlay suppression that involves cortical contrast gain control processes. The current study sought to further explore this notion behaviorally using narrowband and broadband masks, along with a computational neural simulation of the hypothesized underlying gain control processes in cortex. Additionally, we employed transcranial direct current stimulation (tDCS) in order to test whether cortical processes are involved in driving narrowly tuned anisotropic suppression. The behavioral results yielded anisotropic overlay suppression for both broadband and narrowband masks and could be replicated with our computational neural simulation of anisotropic gain control. Further, the anisotropic form of overlay suppression could be directly modulated by tDCS, which would not be expected if the suppression was primarily subcortical in origin. Altogether, the results of the current study provide further evidence in support of an additional overlay suppression process that originates in cortex and show that this form of suppression is also observable with narrowband masks.


Determinants of opioid efficiency in cancer pain: a comprehensive multivariate analysis from a tertiary cancer centre.
Pain is one of the most terrifying symptoms for cancer patients. Although most patients with cancer pain need opioids, complete relief of pain is hard to achieve. This study investigated the factors influencing persistent pain-free survival (PPFS) and opioid efficiency. A prospective study was conducted on 100 patients with cancer pain, hospitalized at the medical oncology clinic of Akdeniz University. Patient records were collected including patient demographics, the disease, treatment characteristics, and details of opioid usage. Pain intensity was measured using a patient self-reported visual analogue scale (VAS). The area under the curve (AUC) reflecting the pain load was calculated from daily VAS tables. PPFS, the primary measure of opioid efficacy, was described as the duration for which a patient reported a greater than or equal to two-point decline in their VAS for pain. Predictors of opioid efficacy were analysed using a multivariate analysis. In the multivariate analysis, PPFS was associated with the AUC for pain (Exp (B)=0.39 (0.23-0.67), P=0.001), the cumulative opioid dosage used during hospitalisation (Exp (B)=1.00(0.99-1.00), P=0.003) and changes in the opioid dosage (Exp (B)=1.01 (1.00-1.01), P=0.016). The change in VAS score over the standard dosage of opioids was strongly associated with current cancer treatment (chemotherapy vs. others) (β=-0.31, T=-2.81, P=0.007) and the VAS for pain at the time of hospitalisation (β=-0.34, T=-3.07, P= 0.003). The pain load, opioid dosage, concurrent usage of chemotherapy and initial pain intensity correlate with the benefit received from opioids in cancer patients.


Differential temporal expression of milk miRNA during the lactation cycle of the marsupial tammar wallaby (Macropus eugenii).
Lactation is a key aspect of mammalian evolution for adaptation of various reproductive strategies along different mammalian lineages. Marsupials, such as tammar wallaby, adopted a short gestation and a relatively long lactation cycle, the newborn is immature at birth and significant development occurs postnatally during lactation. Continuous changes of tammar milk composition may contribute to development and immune protection of pouch young. Here, in order to address the putative contribution of newly identified secretory milk miRNA in these processes, high throughput sequencing of miRNAs collected from tammar milk at different time points of lactation was conducted. A comparative analysis was performed to find distribution of miRNA in milk and blood serum of lactating wallaby. Results showed that high levels of miRNA secreted in milk and allowed the identification of differentially expressed milk miRNAs during the lactation cycle as putative markers of mammary gland activity and functional candidate signals to assist growth and timed development of the young. Comparative analysis of miRNA distribution in milk and blood serum suggests that milk miRNAs are primarily expressed from mammary gland rather than transferred from maternal circulating blood, likely through a new putative exosomal secretory pathway. In contrast, highly expressed milk miRNAs could be detected at significantly higher levels in neonate blood serum in comparison to adult blood, suggesting milk miRNAs may be absorbed through the gut of the young. The function of miRNA in mammary gland development and secretory activity has been proposed, but results from the current study also support a differential role of milk miRNA in regulation of development in the pouch young, revealing a new potential molecular communication between mother and young during mammalian lactation.


Cue Valence Influences the Effects of Cue Uncertainty on ERP Responses to Emotional Events.
Individuals often predict consequences, particularly emotional consequences, according to emotional or non-emotional signals conveyed by environmental cues (i.e., emotional and non-emotional cues, respectively). Some of these cues signify the consequences with certainty (i.e., certain cues), whereas others do not (i.e., uncertain cues). Several event-related potential (ERP) studies regarding non-emotional cues have suggested that the effects of cue uncertainty on attention to emotional events occur in both perception and evaluation processes. However, due to the limitations of previous studies, it is unclear what the effects of cue uncertainty would be in an emotional cue condition. Moreover, it is uncertain whether the effects of cue uncertainty are affected by cue valence (i.e., emotional and non-emotional cues). To address these questions, we asked participants to view cues and then to view emotional (positive or negative) pictures. The cues either did or did not indicate the emotional content of the picture. In the emotional cue condition, happy and fearful faces were used as certain cues indicating upcoming positive and negative pictures, respectively, and neutral faces were used as uncertain cues. In the non-emotional cue condition, scrambled faces outlined in red and blue indicated upcoming positive and negative pictures, respectively, and scrambled faces outlined in green served as uncertain cues. The results showed that for negative pictures, ERP responses in a time range between 60 and 1,000 ms were shifted to a more negative direction in a certain condition than in the uncertain condition when the cues were emotional. However, the effect was the reverse for positive pictures. This effect of cue uncertainty was similar in the non-emotional cue-negative condition. In contrast, there was no effect of cue uncertainty in the non-emotional cue-positive condition. Therefore, the findings indicate that cue uncertainty modulates attention toward emotional events when the events are signified by emotional cues. The findings may also suggest that cue valence modulates the effects of cue uncertainty on attention to emotional events.


Cerebral Palsy for the Pediatric Eye Care Team Part II: Diagnosis and Treatment of Ocular Motor Deficits.
Cerebral palsy is a term used to describe a spectrum of neurological deficits resulting from damage to the developing nervous system. This paper, the second in a series of three articles, will present frequency, diagnosis, and management of the ocular motor deficits associated with CP. Topics for discussion will include the prevalence and type of strabismus, the effect of CP on eye movement systems, the efficacy of vision therapy for eye movement deficits, and strabismus surgery. In 2002, a retrospective chart review of all cases of cerebral palsy referred to the St. Louis Children's Hospital Eye Center was done. Detailed data on the sensory and motor deficits documented in these children was collected. Also recorded was the management strategy and response to treatment. Of the 131 cases reviewed (mean age 5.2 years at presentation), 84 (64%) had strabismus: 55% esotropia and 27% exotropia. Of those patients tested for eye movement disorders 67 (61%) had evidence of instability of fixation; 85% of the patients had a pursuit deficit; and 80% had some type of deficit in performing saccades. Vergence was affected in 45% of our patients. Of the patients who underwent strabismus surgery, 67% achieved success after a mean number of 2.1 surgeries. Surgical success was associated with mild or moderate CP (P <.0005), and a stable preoperative angle. Six percent of our patients underwent vision therapy for strabismus or eye movement anomalies under the direction of an outside optometrist. Infantile strabismus is significantly more common in children with CP than in the general pediatric population. Disorders of eye movements are also very common. Based on our experience and review of the literature there is no convincing evidence that vision therapy significantly improves eye movements or visual functioning. Approximately 2 of every 3 cases of nonaccommodative strabismus associated with CP can be successfully managed with conventional strabismus surgery, though most children will require at least two surgeries to achieve alignment.


Oxyfedrine in myocardial stunning.
Effect of oxyfedrine (OXF)(1 mg/kg) administered just before reperfusion (post-treatment) was investigated in a canine model of myocardial stunning. In the saline-treated animals, myocardial stunning was evidenced by fall in MAP, decrease in LV peak (+) dP/dt, rise in LVEDP and incomplete regeneration of myocardial ATP, after reperfusion. OXF was found to be effective in preventing the haemodynamic and metabolic changes associated with myocardial stunning.


Perceived Versus Actual Aeroallergen Sensitization in Urban Children.
Individuals often report allergy to specific aeroallergens, but allergy testing can reveal disparate sensitization. To characterize the agreement between perceived and actual sensitization to individual aeroallergens in an urban pediatric population. A total of 253 children were enrolled from pediatric clinics in New York, NY. Detailed questionnaires regarding perceived sensitization and serum specific IgE measurements to 10 common aeroallergens were completed. Agreement between perceived and actual sensitization (sIgE ≥ 0.35 kUA/L) to individual aeroallergens was assessed by Cohen's kappa. Multivariable logistic regression models adjusted for potential confounders were used to test for associations between perceived and actual sensitization. A total of 161 (63.6%) of 253 children reported perceived sensitization to 1 or more aeroallergen, and 203 (80.2%) were actually sensitized to 1 or more aeroallergen. Agreement between perceived and actual aeroallergen sensitization was fair for most aeroallergens, with greatest agreement for cat dander (κ, 0.42; 95% CI, 0.32-0.53) and dust (κ, 0.32; 95% CI, 0.20-0.44). Models adjusted for potential confounders showed nearly 6-fold odds of sensitization to cat dander given perceived cat allergy (adjusted odds ratio, 5.82; 95% CI, 2.91-11.64), and over 2-fold odds of sensitization to Dermatophagoides pteronyssinus, Dermatophagoides farinae, dog dander, or grass pollen given perceived sensitization to their respective allergens. Among children with no perceived sensitization, actual sensitization ranged from 5.4% to 30.4%, and was more common for indoor versus outdoor allergens, including cockroach. Children who perceive allergen sensitization to cat, dog, dust, or grass are likely to demonstrate actual sensitization to these individual allergens. Children with no perceived sensitization to allergens are nonetheless frequently sensitized.


Radiomics Can Distinguish Pediatric Supratentorial Embryonal Tumors, High-Grade Gliomas, and Ependymomas.
Pediatric supratentorial tumors such as embryonal tumors, high-grade gliomas, and ependymomas are difficult to distinguish by histopathology and imaging because of overlapping features. We applied machine learning to uncover MR imaging-based radiomics phenotypes that can differentiate these tumor types. Our retrospective cohort of 231 patients from 7 participating institutions had 50 embryonal tumors, 127 high-grade gliomas, and 54 ependymomas. For each tumor volume, we extracted 900 Image Biomarker Standardization Initiative-based PyRadiomics features from T2-weighted and gadolinium-enhanced T1-weighted images. A reduced feature set was obtained by sparse regression analysis and was used as input for 6 candidate classifier models. Training and test sets were randomly allocated from the total cohort in a 75:25 ratio. The final classifier model for embryonal tumor-versus-high-grade gliomas identified 23 features with an area under the curve of 0.98; the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 0.85, 0.91, 0.79, 0.94, and 0.89, respectively. The classifier for embryonal tumor-versus-ependymomas identified 4 features with an area under the curve of 0.82; the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 0.93, 0.69, 0.76, 0.90, and 0.81, respectively. The classifier for high-grade gliomas-versus-ependymomas identified 35 features with an area under the curve of 0.96; the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 0.82, 0.94, 0.82, 0.94, and 0.91, respectively. In this multi-institutional study, we identified distinct radiomic phenotypes that distinguish pediatric supratentorial tumors, high-grade gliomas, and ependymomas with high accuracy. Incorporation of this technique in diagnostic algorithms can improve diagnosis, risk stratification, and treatment planning.


Comparative acute and combinative toxicity of aflatoxin B1 and T-2 toxin in animals and immortalized human cell lines.
Aflatoxin B1 (AFB1) and T-2 toxin (T-2) are important food-borne mycotoxins that have been implicated in human health and as potential biochemical weapons threats. In this study the acute and combinative toxicity of AFB1 and T-2 were tested in F-344 rats, mosquitofish (Gambusia affinis), immortalized human hepatoma cells (HepG2) and human bronchial epithelial cells (BEAS-2B). Preliminary experiments were conducted in order to assess the acute toxicity and to obtain LD50, LC50 and IC50 values for individual toxins in each model, respectively. This was followed by testing combinations of AFB1 and T-2 to obtain LD50, LC50 and IC50 values for the combination in each model. All models demonstrated a significant dose response in the observed parameters to treatment. The potency of the mixture was gauged through the determination of the interaction index metric. The results of this study demonstrate that these two toxins interacted to produce alterations in the toxic responses generally classifiable as additive; however, a synergistic interaction was noted in the case of BEAS-2B. It can be gathered that this combination may pose a significant threat to public health and further research needs to be completed addressing alterations in metabolism and detoxification that may influence the toxic manifestations in combination.


[Surgery for degenerative spondylolisthesis of the lumbar spine using intra-articular fusion. A prospective study].
The aim of the study is to present our surgical method of treating degenerative spondylolisthesis, which includes radical bilateral laminectomy to relieve compression on the spinal cord, transpedicular fixation of the segment and arthrodesis by bilateral intra-articular fusion. This surgery was indicated in patients with grade I or grade II of degenerative sponylolisthesis with a 4-mm or more slippage. Our prospectively studied group consisted of 46 patients (17 men, 29 women; average age, 64.2 years; range, 39-84 years). Before surgery and at 1 year after the procedure, the intensity of axial pain and that of radicular pain were each assessed using the visual Analogue Scale (VAS). Difficulty in performing daily living activities was measured by the Oswestry Disability Index (ODI). The surgical procedure included laminectomy, partial medial facetectomy, foraminotomy to relieve pressure on the spinal nerve roots and transpedicular fixation to provide stability. Using a cutter, cartilage was separated off the cortical bone and, in order to facilitate fusion, bone cavities thus produced were filed with corticospongious grafts harvested from the removed vertebral arch with Kerrison forceps. At 1-year follow-up, dynamic X-ray was used to evaluate spine alignment and, on a CT scan, the degree of intra-articular fusion was assessed. Fusion was achieved when bone density measurement showed more than 350 Hounsfield Units (HU). For the measurements, the authors used their own modified method by means of a Region of Interest (ROI) analysis. The clinical and radiographic results were statistically evaluated. At 1 year after surgery, lumbar flexion-extension bending X-ray films revealed stability of the treated segments in all patients (100%). CT examination showed bone density higher than 350 HU at both joints, i.e., complete bone fusion, also in all 46 patients. The mean post-operative ODI score was significantly lower than its mean pre-operative value (23.6 vs 55.4), which was improvement by 57.4%. The differences in pre- and post-operative VAS scores were also statistically significant. The mean VAS score for low back pain decreased from 7.61 to 1.74, i.e., improvement by 77.1%, and the mean vAS score for radicular pain dropped from 6.98 to 1.24, i.e., improvement by 82.2%. Assessed by Odom's outcome criteria, the results were excellent in 26 patients and very good in 20 patients, and they were not related to age, gender or the spinal level treated. Any complications associated with the operative procedure or wound healing and requiring repeated surgical treatment were not recorded. The surgical technique described here has advantages over other methods in reliable achieving nerve decompression, joint fusion and spinal stability at low costs and short operative time. In addition, it avoids the necessity of harvesting bone from the iliac crest. At 1-year follow-up all patients showed better health conditions, with improvement in average scores for the ODI by 57%, for low back pain by 77% and for radicular and claudication pain by 82%. The technique of intra-articular fusion for treatment of degenerative spondylolisthesis resulted in solid bone fusion and spinal stability in all patients.


Respiratory plasticity is insufficient to alleviate blood acid-base disturbances after acclimation to ocean acidification in the estuarine red drum, Sciaenops ocellatus.
The changes in ocean chemistry stemming from anthropogenic CO2 release--termed ocean acidification (OA)--are predicted to have wide-ranging effects on fish and ultimately threaten global populations. The ability of fish to adapt to environmental change is currently unknown, but phenotypic plasticity has been highlighted as a crucial factor in determining species resilience. Here we show that red drum, a long-lived estuarine-dependent fish species native to the Gulf of Mexico, exhibit respiratory plasticity that increases CO2 excretion capacity when acclimated to OA conditions. Specifically, fish exposed to 14 days of 1000 µatm CO2 had a 32% reduction in branchial diffusion distance and increased expression of two putative CO2 channel proteins--rhag and rhcg1. No changes were observed in the erythrocyte CO2 transport pathways. Surprisingly, no significant changes in blood chemistry were observed between acclimated and acutely challenged animals; however, a non-significant 30 % drop in the magnitude of plasma C(CO2) elevation was observed. Reduced diffusion distance also comes with the cost of increased diffusive water loss, which would require greater osmoregulatory investment by the animal. OA exposure induced increased gill Na(+), K(+) ATPase activity and intestinal nkcc2 expression, supporting both the presumed osmotic stress and increased osmoregulatory investment. However, no differences in standard metabolic rate, maximum metabolic rate or aerobic scope were detected between control and OA acclimated individuals. Similarly, no differences in critical swim speed were detected between groups, suggesting the energetic cost related to respiratory plasticity is negligible against background metabolism. The current study demonstrated that red drum exhibit respiratory plasticity with only mild physiological trade-offs; however, this plasticity is insufficient to fully offset the OA-induced acid-base disturbance and as such is unlikely to impact species resilience.


Purification of matrix metalloproteinases by column chromatography.
Matrix metalloproteinases (MMPs) are zinc endopeptidases composed of 23 members in humans, which belong to a subfamily of the metzincin superfamily. They play important roles in many pathophysiological events including development, organogenesis, angiogenesis, tissue remodeling and destruction, and cancer cell proliferation and progression by degradation of extracellular matrix (ECM) and non-ECM proteins and interaction with various molecules. Here, we present standard protocols for purification of native proMMPs (proMMP-1, -2, -3, -7, -9 and -10) and recombinant MT1-MMP (MMP-14) using conventional column chromatography. Purification steps comprise the initial common step [diethylaminoethyl (DEAE)-cellulose, Green A Dyematrex gel and gelatin-Sepharose columns], the second step for removal of nontarget proMMPs by immunoaffinity columns (anti-MMP-1 and/or anti-MMP-3 IgG-Sepharose columns) and the final step for further purification (IgG-Sepharose, DEAE-cellulose, Zn2+-chelate-Sepharose and/or gel filtration columns). Purified proMMPs and MMP are functionally active and suitable for biochemical analyses. The basic protocol for the purification from culture media takes approximately 7-10 d.


Disseminated intravascular coagulation in a patient with progressive systemic sclerosis associated with necrotizing angiitis and generalized lymphadenopathy.
A 47-year-old woman with progressive systemic sclerosis developed disseminated intravascular coagulation in the course of her terminal illness. She also had complicating necrotizing angiitis and generalized lymphadenopathy. The likely relationship between disseminated intravascular coagulation and vasculitis is discussed.


Carbon nanotube-DNA nanoarchitectures and electronic functionality.
Biological molecules such as deoxyribonucleic acid (DNA) possess inherent recognition and self-assembly capabilities, and are attractive templates for constructing functional hierarchical material structures as building blocks for nanoelectronics. Here we report the assembly and electronic functionality of nanoarchitectures based on conjugates of single-walled carbon nanotubes (SWNTs) functionalized with carboxylic groups and single-stranded DNA (ssDNA) sequences possessing terminal amino groups on both ends, hybridized together through amide linkages by adopting a straightforward synthetic route. Morphological and chemical-functional characterization of the nanoarchitectures are investigated using scanning electron microscopy, transmission electron microscopy, atomic force microscopy, energy-dispersive X-ray spectroscopy, Raman spectroscopy, and Fourier-transform infrared spectroscopy. Electrical measurements (I-V characterization) of the nanoarchitectures demonstrate negative differential resistance in the presence of SWNT/ssDNA interfaces, which indicates a biomimetic route to fabricating resonant tunneling diodes. I-V characterization on platinum-metallized SWNT-ssDNA nanoarchitectures via salt reduction indicates modulation of their electrical properties, with effects ranging from those of a resonant tunneling diode to a resistor, depending on the amount of metallization. Electron transport through the nanoarchitectures has been analyzed by density functional theory calculations. Our studies illustrate the great promise of biomimetic assembly of functional nanosystems based on biotemplated materials and present new avenues toward exciting future opportunities in nanoelectronics and nanobiotechnology.


Heterologous expression and epitope mapping of a human small nuclear ribonucleoprotein-associated Sm-B'/B autoantigen.
The U snRNP associated B'/B polypeptides are primary targets of Sm autoantibodies in patients with systemic lupus erythematosus. We have bacterially expressed a Sm-B'/B autoantigen from Raji cells as a fusion with the anthranilate synthase protein from Escherichia coli. The recombinant Sm-B'/B fusion displays comparable immunologic reactivity to the native protein when tested with both monoclonal and polyclonal antibodies. To map Sm-B'/B epitopes, we constructed a series of 12 anthranilate synthase fusions spanning different regions of Sm-B'/B and tested such fusions on immunoblots against a panel of characterized sera. In this manner, we have identified six epitopes, five of which overlap the proline-rich carboxyl-terminus of the protein. Some of these epitopes appear to be conformational. The human sera tested can be divided, according to the epitopes they recognize, into six groups. Finally, we have shown that anti-Sm recognition of the (U1)RNP-specific A protein is attributable to cross-reactivity between the Sm-B'/B and A autoantigens.


Patient and clinician experiences with telehealth for patient follow-up care.
The increasing and widespread availability of personal technology offers patients and clinicians the opportunity to utilize real-time virtual communication to enhance access to health services. Understanding the perceived value of different modes of care may help to shape the future use of technology. Cross-sectional surveys of patients and clinicians participating in telehealth virtual video visits (VVVs) in an academic health system. We administered surveys to 426 unique established patients and 74 attending physicians in our hospital to measure perceptions of the comparative experience of VVVs and office visits; 254 patients and 61 physicians completed the surveys. When comparing VVVs and office visits, 62.6% of patients and 59.0% of clinicians reported no difference in "the overall quality of the visit." VVVs were vastly preferred to office visits by patients for convenience and travel time. A majority (52.5%) of clinicians reported higher efficiency of a VVV appointment. For established patients, VVVs may provide effective follow-up and enhanced convenience when compared with traditional office visits.


In Vivo Capsid Engineering of Bacteriophages for Oriented Surface Conjugation.
The current state-of-the-art in bacteriophage (phage) immobilization onto magnetic particles is limited to techniques that are less expensive and/or facile but nonspecific or those that are more expensive and/or complicated but ensure capsid-down orientation of the phages, as necessary to preserve infectivity and performance in subsequent applications (e.g., therapeutics, detection). These cost, complexity, and effectiveness limitations constitute the major hurdles that limit the scale-up of phage-based strategies and thus their accessibility in low-resource settings. Here, we report a plasmid-based technique that incorporates a silica-binding protein, L2, into the T7 phage capsid, during viral assembly, with and without inclusion of a flexible linker peptide, allowing for targeted binding of the phage capsid to silica without requiring the direct modification of the phage genome. L2-tagged phages were then immobilized onto silica-coated magnetic nanoparticles. Inclusion of the flexible linker between the phage capsid protein and the L2 protein improved immobilization density compared to both wild type T7 phages and L2-tagged phages without the flexible linker. Taken together, this work demonstrates phage capsid modification without engineering the phage genome, which provides an important step toward reducing the cost and increasing the specificity/directionality of phage immobilization methods and could be more broadly applied in the future for other phages for a range of other capsid tags and nanomaterials.


Serum antibody levels to the Pneumocystis jirovecii major surface glycoprotein in the diagnosis of P. jirovecii pneumonia in HIV+ patients.
Pneumocystis jirovecii remains an important cause of fatal pneumonia (Pneumocystis pneumonia or PcP) in HIV+ patients and other immunocompromised hosts. Despite many previous attempts, a clinically useful serologic test for P. jirovecii infection has never been developed. We analyzed serum antibody responses to the P. jirovecii major surface glycoprotein recombinant fragment C1 (MsgC1) in 110 HIV+ patients with active PcP (cases) and 63 HIV+ patients with pneumonia due to other causes (controls) by an enzyme-linked immunosorbent assay (ELISA). The cases had significantly higher IgG and IgM antibody levels to MsgC1 than the controls at hospital admission (week 0) and intervals up to at least 1 month thereafter. The sensitivity, specificity and positive predictive value (PPV) of IgG antibody levels increased from 57.2%, 61.7% and 71.5% at week 0 to 63.4%, 100%, and 100%, respectively, at weeks 3-4. The sensitivity, specificity and PPV of IgM antibody levels rose from 59.7%, 61.3%, and 79.3% at week 0 to 74.6%, 73.7%, and 89.8%, respectively, at weeks 3-4. Multivariate analysis revealed that a diagnosis of PcP was the only independent predictor of high IgG and IgM antibody levels to MsgC1. A high LDH level, a nonspecific marker of lung damage, was an independent predictor of low IgG antibody levels to MsgC1. The results suggest that the ELISA shows promise as an aid to the diagnosis of PCP in situations where diagnostic procedures cannot be performed. Further studies in other patient populations are needed to better define the usefulness of this serologic test.


Isolation and Analysis of hupR Gene Required for the Expression of Hydrogenase in Rhodobacter sphaeroides.
Cosmid 1 containing the hup genes isolated from the photosynthetic bacterium Rhodobacter sphaeroides was studied. The hupR gene from cosmid 1 was cloned and sequenced (EMBL accession number AJ243734). It encoded a 54.031 kD protein homologous to transcriptional regulators belonging to the superfamily of two-component regulatory systems. The HupR protein was overexpressed in Escherichia coli in the form of His6-tagged HupR. The cloned hupR gene could restore hydrogenase activity in R.sphaeroides hupR mutants and activate hupSL gene transcription.


Structure-function relations in photosystem II. Effects of temperature and chaotropic agents on the period four oscillation of flash-induced oxygen evolution.
The characteristic period four oscillation patterns of oxygen evolution induced by a train of single-turnover flashes were measured in dark-adapted samples as a function of temperature and upon addition of chaotropic agents. The following results were obtained: (a) Within the range of 0 < theta < 35 degrees C, the ratio of the oxygen yield induced by the 4th and 3rd flashes of the train, Y4/Y3, and the oxygen yield induced by the 2nd flash, Y2, exhibit similar dependencies on the temperature in isolated thylakoids, PS II membrane fragments, and inside-out vesicles. (b) Below a characteristic temperature theta c of 20-25 degrees C, the values of Y4/Y3 and Y2, which reflect (at constant S0 dark population) the probabilities of misses and double hits, respectively, remain virtually independent of temperature, whereas above theta c these parameters increase. (c) The dark decays of S2 and S3 via fast and slow kinetics due to reduction of the water oxidase by YD and other endogenous electron donor(s), respectively, exhibit comparatively strong temperature dependencies in thylakoids with the following activation energies: EA(S2fast) = 55 kJ/mol, EA(S3fast) = 50 kJ/mol, EA(S2slow) = 85 kJ/mol, and EA(S3slow) = 75 kJ/mol. The activation energy of S0 oxidation to S1 by YDox was found to be markedly smaller with a value of EA(S0) = 30 kJ/mol. (d) Incubation with chaotropic agents at concentrations which do not significantly impair the oxygen evolution capacity leads to modifications of the oscillation pattern with remarkable differences for various types of agents: Tris and urea are practically without effect; guanidine hydrochloride affects Y4/Y3 in a similar way as elevated temperature but without significant changes of Y2 and the decay kinetics of S2 and S3; and anions of the Hofmeister series (SCN-, ClO4-, I-) cause a drastic destabilization of YDox. Possible structure-function relations of the PS II complex are discussed on the basis of these findings.


Dexamethasone inhibits feedback regulation of the mitogenic activity of tumor necrosis factor, interleukin-1, and epidermal growth factor in human fibroblasts.
Tumor necrosis factor (TNF), interleukin-1 (IL-1), and epidermal growth factor (EGF) were mitogenic for human diploid FS-4 fibroblasts. Dexamethasone amplified the growth-stimulating action of all three agents. Amplification of the growth-stimulating action was maximal when dexamethasone was added along with TNF or EGF; no amplification was seen if the addition of dexamethasone was delayed for more than 3 hr. Prolonged simultaneous treatment with TNF and EGF resulted in less growth stimulation than treatment with EGF alone. Dexamethasone abolished this apparent antagonistic interaction between TNF and EGF. Dexamethasone also inhibited the antiviral action of TNF against encephalomyocarditis (EMC) virus in FS-4 cells. TNF and IL-1 increased the steady state level of interferon (IFN)-beta 2 mRNA but failed to induce detectable levels of IFN-beta 1 mRNA in FS-4 cells. Dexamethasone inhibited the increase of IFN-beta 2 mRNA levels by IL-1 or TNF. Inhibition of IFN-beta synthesis is likely to be responsible for the inhibition of the TNF-induced antiviral state by dexamethasone. Since IFNs suppress cell growth, inhibition of endogenous IFN-beta synthesis may also be responsible for the amplification by dexamethasone of the growth-stimulating action of TNF and IL-1. Amplification of the mitogenic action of EGF by dexamethasone appears to be mediated by different mechanism.


The Mutual Relationship Between Men's Drinking and Depression: A 4-Year Longitudinal Analysis.
The purpose of the current study was to examine the longitudinal reciprocal relationship between depression and drinking among male adults from the general population. This study used a panel dataset from the Korean Welfare Panel (from 2011 to 2014). The subjects were 2511 male adults aged between 20 and 65 years. Based on the Korean Version of the Alcohol Use Disorders Identification Test (AUDIT-K) scores, 2191 subjects were categorized as the control group (AUDIT-K < 12) and 320 subjects were categorized as the problem drinking group (AUDIT-K ≥ 12). An autoregressive cross-lagged modelling analysis was performed to investigate the mutual relationship between problem drinking and depression measured consecutively over time. The results indicated that alcohol drinking and depression were stable over time. In the control group, there was no significant causal relationship between problem drinking and depression while in the problem drinking group, drinking in the previous year significantly influenced depression in the following second, third and fourth years. This study compared normal versus problem drinkers and showed a 4-year mutual causal relationship between depression and drinking. No longitudinal interaction between drinking and depression occurred in normal drinkers, while drinking intensified depression over time in problem drinkers. This study found that problem drinking was a risk factor for development of depression. Therefore, more attention should be given to problem alcohol use in the general population and evaluation of past alcohol use history in patients with depressive disorders.


The parathyroid adenoma: an imaging/surgical perspective.
The clinical picture of hyperparathyroidism has changed since the implementation of routine serum calcium testing, resulting in more asymptomatic patients undergoing early surgical exploration. Although operative complications (e.g., recurrent laryngeal nerve paralysis, hypocalcemia, etc.) are not prevalent, the risk can be minimized by minimizing tissue dissection. For this reason, we feel that preoperative tumor localization is of great importance. We report our imaging results of parathyroid adenomas, utilizing ultrasonography, technetium-thallium subtraction scanning, digital subtraction angiography and magnetic resonance imaging. We also present an imaging protocol which, we have found, maximizes preoperative identification of these tumors.


Structure and transcriptional regulation of the gene encoding pyruvate formate-lyase of a ruminal bacterium, Streptococcus bovis.
The gene (pfl) encoding pyruvate formate-lyase (Pfl) from Streptococcus bovis was sequenced. The deduced amino acid sequence of Pfl was similar to Streptococcus mutans Pfl, and included the conserved regions necessary for free-radical formation and a catalytic site. The Pfl of S. bovis appeared to be a free-radical-containing enzyme because of its dioxygen sensitivity and its amino acid sequence similarity with the Escherichia coli enzyme. The pfl mRNA of S. bovis was approximately 2.3 kb and was transcribed in a monocistronic fashion. When cells were grown in batch culture at pH 6.9, the level of pfl transcript increased as the growth phase changed from exponential growth to stationary phase. This result was in constrast to the previous observation that the level of lactate dehydrogenase (Ldh) mRNA decreased during the later stages of growth. Continuous culture experiments conducted at pH 6.9 under glucose-limited and ammonia-limited conditions revealed that pfl mRNA was decreased by an excess supply of glucose, as well as by a high growth rate. On the contrary, ldh mRNA increased when excess glucose was supplied and the growth rate was high. The amount of pfl mRNA in cells was lower at pH 4.5 than pH 6.9, whereas the level of ldh mRNA was higher at pH 4.5. This result was consistent with the amounts of Pfl and Ldh in cells and the proportion of formate and lactate produced. These results support the hypothesis that S. bovis regulates Pfl and Ldh synthesis at the transcriptional level in response to growth conditions.


Phosphorylation of nonstructural 5A protein of hepatitis C virus: HCV group-specific hyperphosphorylation.
We previously showed that two proteins with molecular weights of 56 and 58 kDa are produced from nonstructural protein 5A (NS5A) derived from hepatitis C virus (HCV)-1b genotype. The 56-kDa protein is phosphorylated at serine residues in NS5A, including those located in the C-terminal region of NS5A, while the 58-kDa protein, the hyperphosphorylated form of the 56-kDa protein, is phosphorylated at serine residues in the central region. This hyperphosphorylation is dependent on the presence of HCV NS4A protein. To clarify whether NS4A-dependent phosphorylation also occurs in other HCV genotypes, phosphorylation of NS5A was analyzed by two-dimensional gel electrophoresis. Here, we report that NS5A from the HCV-2a genotype was phosphorylated. However, hyperphosphorylation of NS5A occurs in the HCV-1b genotype but not in the -2a genotype. This result suggests that modification of NS5A phosphorylation reflects the virological features of HCV and that there are physiological differences in the roles of differently phosphorylated NS5A between HCV genotypes.


Effect of Pd and In on mercury evaporation from amalgams.
The amount of Hg vapor released from "synthesized" gamma 1 with 1% (wt) Pd was reported to be less than 30% of that from gamma 1 with no Pd. This study tested the hypothesis that Hg evaporation from Pd-containing amalgams decreases with Pd concentration and that In also reduces Hg vapor. Specimens (4 mm dia, 8 mm long) were prepared by triturating Ag-Sn(25%)-Cu(12%) alloy powder containing 0.5-9.0% Pd with pure Hg and by triturating 3% Pd alloy powder with Hg containing 1-5% In (all residual Hg approximately equal to 62%). The total amount (ng/mm2) of Hg vapor released at 37 degrees C from freshly prepared amalgams was measured. Pd (3-9%) in the powder significantly (p < 0.05) decreased Hg vapor release from amalgams during setting. Use of In-containing Hg also reduced Hg vapor release (5% In, p < 0.05). The reduction in the Hg vapor pressure by adding Pd and the rapid oxide film formation on the In-containing amalgam appear to work together to reduce Hg vaporization from these amalgams.


The expression of low density lipoprotein receptor-related protein in colorectal carcinoma.
Low density lipoprotein receptor-related protein (LRP) is a multifunctional cell surface receptor binding many different ligands including proteinases and their inhibitors, some of which are known to be involved in tumor biology. We studied the expression of LRP and its putative role in colorectal carcinoma. Tissue samples were obtained from 50 patients with colorectal carcinoma and fixed in formalin and embedded in paraffin. Immunohistochemical staining was performed using antibodies directed against LRP, cathepsin B and urokinase-type plasminogen activator (u-PA). The expression of LRP was further studied by polymerase chain reaction. The TNM stage was determined according to UICC guide lines and was based upon histological analysis. LRP was primarily expressed in stroma cells [36 patients (72%)] and less frequently in tumor cells [6 patients (12%)]. In 22% of all cases LRP was prominent at the invasion front. Cathepsin B was found both in the tumor stroma [50 (100%)] and in the tumor cells [46 (92%)]. u-PA was present in the tumor stroma [44 (88%)] and in the tumor cells [44 (88%)]. In stromal cells the expression of LRP correlated significantly with the expression of u-PA (p=0.043). Furthermore, the expression of LRP and of u-PA in tumor cells correlated with the tumor stage according to UICC (p=0.038 and 0.018, respectively). We provide evidence that LRP is expressed in colorectal cancer. As LRP forms complexes with u-PA and its inhibitor, we suspect that LRP can influence the known effects of u-PA on tumor biology.


The bilateral strength and power asymmetries in untrained boys.
The aim of the study was to identify the level of isokinetic strength and power of lower limbs in 13-year-old untrained boys (n=22, height: 158.5±8.0 cm, mass: 49.1±12.6 kg), to determine bilateral deficit between the limbs in the tests and examine their mutual relationship. Maximum peak muscle torque of knee extensors (PTQ) and flexors (PTH) on dominant (DL) and non-dominant leg (NL) were measured by isokinetic dynamometer. Three types of a vertical jump: countermovement jump with (CMJFA) and without arms (CMJ) and squat jump (SJ) were performed on two force platforms. We found the significant effect (p<.01) of independent variables (knee extensors, flexors, AV) and their interaction on PT. AV did not indicate any significant effect on bilateral ratio of knee extensors (F4,84=.74, p>.05, ηp2=0.03), however a significant effect of AV was found in knee flexors (F4,84=2.70, p<.05, ηp2=.114). The type of jump had no effect on the difference between force exerted by DL and NL (F1,21=.102, p>.05, ηp2=.01). Bilateral deficit (Q:Q, H:H) did not significantly correlate with bilateral deficit in jumps (p>.05). Despite the possibility of identifying muscle asymmetries in the sense of strength imbalances, their mutual relationship with results in isokinetic dynamometry and power jump tests is still unclear.


Impact of rainfall properties on the performance of hydrological models for green roofs simulation.
Green roofs (GR) are effective tools for the mitigation of the negative hydrological impact linked to uncontrolled urbanization. Models for runoff response of vegetated covers support planning decisions about the use of this technology in ever-expanding areas, but there is still large uncertainty in this research area. The goal of the present study was to define the accuracy of three selected models for the simulation of the hydrological behavior of a GR, with a particular focus on the precipitation pattern characteristics. The Nash cascade model, Storm Water Management Model (SWMM), and HYDRUS-1D models were selected. Each model was calibrated over 24 rainfall-runoff events collected at the experimental site located in Southern Italy. Rainfall events were characterized using several criteria and were divided into convective, stratiform, and tropical precipitation types according to the shape of the rainfall profile identified by the rainfall binary shape code. The models returned overall satisfactory performances with average Nash-Sutcliffe Efficiency index of 0.65 for the SWMM and HYDRUS and 0.73 for the Nash model. In general, the models were better performing for stratiform and tropical events. SWMM and HYDRUS predicted with higher accuracy the convective events while the Nash model appeared more suitable for stratiform events.


ISLES 2015 - A public evaluation benchmark for ischemic stroke lesion segmentation from multispectral MRI.
Ischemic stroke is the most common cerebrovascular disease, and its diagnosis, treatment, and study relies on non-invasive imaging. Algorithms for stroke lesion segmentation from magnetic resonance imaging (MRI) volumes are intensely researched, but the reported results are largely incomparable due to different datasets and evaluation schemes. We approached this urgent problem of comparability with the Ischemic Stroke Lesion Segmentation (ISLES) challenge organized in conjunction with the MICCAI 2015 conference. In this paper we propose a common evaluation framework, describe the publicly available datasets, and present the results of the two sub-challenges: Sub-Acute Stroke Lesion Segmentation (SISS) and Stroke Perfusion Estimation (SPES). A total of 16 research groups participated with a wide range of state-of-the-art automatic segmentation algorithms. A thorough analysis of the obtained data enables a critical evaluation of the current state-of-the-art, recommendations for further developments, and the identification of remaining challenges. The segmentation of acute perfusion lesions addressed in SPES was found to be feasible. However, algorithms applied to sub-acute lesion segmentation in SISS still lack accuracy. Overall, no algorithmic characteristic of any method was found to perform superior to the others. Instead, the characteristics of stroke lesion appearances, their evolution, and the observed challenges should be studied in detail. The annotated ISLES image datasets continue to be publicly available through an online evaluation system to serve as an ongoing benchmarking resource (www.isles-challenge.org).


Basis of tetrodotoxin's selectivity in blockage of squid axons.
The blockage of nerve activity by tetrodotoxin is unusually potent and specific. Our experiments were designed to distinguish whether its specificity of action was based on the identification of ions, the direction of cation flow, or differences in the early transient and late steady conductance pathways. Alkali cations were substituted for sodium in the sea water, bathing an "artificial node" in a voltage-clamped squid axon. When tetrodotoxin was added to the artificial sea waters at a concentration of 100 to 150 mM, it was found to always block the flow of cations through the early transient channel, both inward and outward, but it never blocked the flow of ions using the late steady pathway. We conclude that the selectivity of tetrodotoxin is based on some difference in these two channels.


Design and operation of a fast electromagnetic inductive massive gas injection valve for NSTX-U.
Results from the operation of an electromagnetic valve, that does not incorporate ferromagnetic materials, are presented. Image currents induced on a conducting disc placed near a pancake solenoid cause it to move away from the solenoid and open the vacuum seal. A new and important design feature is the use of Lip Seals for the sliding piston. The pressure rise in the test chamber is measured directly using a fast time response Baratron gauge. The valve injects over 200 Torr l of nitrogen in less than 3 ms, which remains unchanged at moderate magnetic fields.


Solutions for vectorial beam coupling under ac field in cubic photorefractive crystals.
We develop a theory of vectorial wave coupling in cubic photorefractive crystals placed in an alternating ac field to enhance the nonlinear response. It is proven in the general case that despite essential differences between the diffusion and the ac nonlocal responses, the latter keeps the light interference fringes straight during the interaction. This fundamental feature allows, under certain restrictions, to reduce the nonlinear problem of vectorial coupling to the known linear problem of vectorial Bragg diffraction from a spatially uniform grating, which admits an exact solution. As a result, the nonlinear vectorial problem can be effectively solved for a number of practically important cases. These cases include nontrivial polarization effects and also the high-contrast effects. The presence of conservation laws involving the polarization degrees of freedom is shown. A number of particular cases relevant to experiments with BTO crystals are considered.


Unlikely Fluctuations and Non-Equilibrium Work Theorems-A Simple Example.
An exciting development in statistical mechanics has been the elucidation of a series of surprising equalities involving the work done during a nonequilibrium process. Astumian has presented an elegant example of such an equality, involving a colloidal particle undergoing Brownian motion in the presence of gravity. We analyze this example; its simplicity, and its link to geometric Brownian motion, allows us to clarify the inner workings of the equality. Our analysis explicitly shows the important role played by large, unlikely fluctuations.


[Syncopes in patients of advanced age].
This review covers the various etiologies and therapeutic possibilities of syncopes in the elderly. The importance of normal blood pressure and the danger of hypotension are stressed: The frequently occurring vascular stenoses may become especially dangerous if pre-stenotic pressure drops as a consequence of orthostatic dysregulation or of inadequate treatment of hypertension. Orthostatic dysregulation--a common phenomenon in old age--is possibly due to a diminished activity of the baroreceptor reflexes or to the pooling of blood in the venous system and/or to the diminution of blood volume by therapeutic procedures (diuretics) or by bed rest. The cautious use of antihypertonics, of diuretics and of psychopharmacological drugs in the correctly adjusted dose is essential in the management of these pathophysiological mechanisms in the elderly. The orthostatic hypotension--a condition that has to be treated in the old aged--can be managed by supportive treatment such as education of baroreceptor reflexes, compression of the lower extremities, and correction of electrolyte and water disturbances. Vasoconstrictors have to be used additionally with great care to adjust the correct dose of the combination of dihydroergotamine and sympathomimetics individually.


Decision analysis tree for deciding whether to remove an undescended testis from a young dog.
A decision analysis tree was constructed to estimate the life span which might be expected for a dog presented with cryptorchidism at one year of age if it underwent a preventive orchidectomy or if it did not. The tree was constructed by using risk factors associated with cryptorchidism and data on the prevalence of complications after surgery from the literature. The expected life span without an orchidectomy was not significantly different from the expected life span after an orchidectomy.


Hyperparasitemia and blood exchange transfusion for treatment of children with falciparum malaria.
A substantial number of reports of cases and small investigations support blood exchange transfusion as a therapy for hyperparasitemia in cases of Plasmodium falciparum malaria, although a prospective and randomized study has never been undertaken. We report on 113 prospectively enrolled children in Lambaréné, Gabon, who had more than 10% parasitized erythrocytes and were treated with chemotherapy alone. All 86 patients with hyperparasitemia as the sole complication recovered uneventfully. Among the 27 patients who had additional complications, parasitemia levels as high as 81% responded well to chemotherapy alone. Two patients with cerebral malaria, who also had other complications and hence a poor prognosis, died. Hyperparasitemia itself might be important for the development of a fatal event in malaria, but a recommendation to perform a dangerous, expensive, and labor-intensive procedure such as blood exchange transfusion for its treatment should be based on substantial clinical research, especially in areas where malaria is a major health problem.


[Not Available].
This essay focuses on the Augsburger barber-surgeon Joseph Schmid (1601/1606(?)-1667), a typical representative of his guild in regard to his apprenticeship and profession. But his surgical publications - written for a wide public as well as for future barber-surgeons - set him apart from the mass of barber-surgeons. His publications became well-known in the 17th and early 18th century and they belonged in "a good German surgical library". In his book entitled "Speculum Chirurgicum", published in 1656, he gave an insight into his patients and profession.


Group A Streptococcal Pericarditis in a Four-Month-Old Infant: Case report.
Purulent pericarditis is uncommon among paediatric patients and cases caused by group A Streptococcus (GAS) are even rarer. We report a four-month-old female infant who was referred to the Royal Hospital, Muscat, Oman, in 2015 with pericardial effusion and cardiac tamponade. She had initially presented to a secondary hospital with a two-week history of fever, a runny nose and shortness of breath. Blood and pericardial fluid cultures confirmed GAS isolates. The infant was treated with a two-week course of antibiotics and made a complete recovery with no echocardiographical evidence of pericardial effusion at a two-month follow-up. To the best of the authors' knowledge, this case constitutes the youngest infant to present with GAS pericarditis. As invasive GAS infections can present in infancy, early recognition and treatment is required.


Preclinical ex-vivo Testing of Anti-inflammatory Drugs in a Bovine Intervertebral Degenerative Disc Model.
Discogenic low back pain (LBP) is a main cause of disability and inflammation is presumed to be a major driver of symptomatic intervertebral disc degeneration (IDD). Anti-inflammatory agents are currently under investigation as they demonstrated to alleviate symptoms in patients having IDD. However, their underlying anti-inflammatory and regenerative activity is poorly explored. The present study sought to investigate the potential of Etanercept and Tofacitinib for maintaining disc homeostasis in a preclinical intervertebral disc (IVD) organ culture model within IVD bioreactors allowing for dynamic loading and nutrient exchange. Bovine caudal IVDs were cultured in a bioreactor system for 4 days to simulate physiological or degenerative conditions: (1) Phy-physiological loading (0.02-0.2 MPa; 0.2 Hz; 2 h/day) and high glucose DMEM medium (4.5 g/L); (2) Deg+Tumor necrosis factor α (TNF-α)-degenerative loading (0.32-0.5 MPa; 5 Hz; 2 h/day) and low glucose DMEM medium (2 g/L), with TNF-α injection. Etanercept was injected intradiscally while Tofacitinib was supplemented into the culture medium. Gene expression in the IVD tissue was measured by RT-qPCR. Release of nitric oxide (NO), interleukin 8 (IL-8) and glycosaminoglycan (GAG) into the IVD conditioned medium were analyzed. Cell viability in the IVD was assessed using lactate dehydrogenase and ethidium homodimer-1 staining. Immunohistochemistry was performed to assess protein expression of IL-1β, IL-6, IL-8, and collagen type II in the IVD tissue. Etanercept and Tofacitinib downregulated the expression of IL-1β, IL-6, IL-8, Matrix metalloproteinase 1 (MMP1), and MMP3 in the nucleus pulposus (NP) tissue and IL-1β, MMP3, Cyclooxygenase-2 (COX2), and Nerve growth factor (NGF) in the annulus fibrosus (AF) tissue. Furthermore, Etanercept significantly reduced the IL-1β positively stained cells in the outer AF and NP regions. Tofacitinib significantly reduced IL-1β and IL-8 positively stained cells in the inner AF region. Both, Etanercept and Tofacitinib reduced the GAG loss to the level under physiological culture condition. Etanercept and Tofacitinib are able to neutralize the proinflammatory and catabolic environment in the IDD organ culture model. However, combined anti-inflammatory and anabolic treatment may be required to constrain accelerated IDD and relieving inflammation-induced back pain.


Fog dispersal by CO(2) laser pulses: an exact solution including prevaporization heating.
The coupled nonlinear equations governing the transmission of a collimated high-energy laser beam through a fog are solved analytically. The incident radiation is assumed to be of sufficient intensity to vaporize the droplets in the path of the beams, thus enabling a greater percentage than without vaporization of the incident energy to be transmitted. Prevaporization heating is included in the treatment, and the vaporization front provides a moving boundary condition for the solution. The initial laser intensity and aerosol density are taken to be arbitrary functions of time and position, respectively; Effects of scattering, molecular absorption, and heat conduction are ignored.


Proteolytic processing of human factor VIII. Correlation of specific cleavages by thrombin, factor Xa, and activated protein C with activation and inactivation of factor VIII coagulant activity.
Human factor VIII was isolated from commercial factor VIII concentrates and found to consist of multiple polypeptides with molecular weights ranging from 80 000 to 210 000. Immunological and amino acid sequence data identified these polypeptides as subunits of factor VIII. N-Terminal amino acid sequence analysis determined that the Mr 210 000 and 80 000 proteins are derived from the N- and C-terminal portions of factor VIII, respectively; Mr 90 000-180 000 polypeptides are derived from the Mr 210 000 polypeptide by C-terminal cleavages. Treatment of purified factor VIII with thrombin resulted in proteolysis of Mr 80 000-210 000 proteins and the generation of polypeptides of Mr 73 000, 50 000, and 43 000. Maximum coagulant activity of thrombin-activated factor VIII was correlated with the generation of these polypeptides. The proteolysis as well as activation of factor VIII by thrombin was found to be markedly dependent on CaCl2 concentration. Proteolysis of factor VIII with activated protein C (APC) resulted in degradation of the Mr 90 000-210 000 proteins with the generation of an Mr 45 000 fragment. This cleavage correlated with inactivation of factor VIII by APC. The Mr 80 000 protein was not degraded by APC. Factor Xa cleaved the Mr 80 000-210 000 factor VIII proteins, resulting in the generation of fragments of Mr 73 000, 67 000, 50 000, 45 000, and 43 000. Factor Xa was found to initially activate and subsequently inactivate factor VIII.(ABSTRACT TRUNCATED AT 250 WORDS)


Reassembled plasma low density lipoproteins. Phospholipid-cholesterol ester-apoprotein B complexes.
Reassembled low density lipoprotein (LDL) complexes have been prepared by the interaction of lipid-free sodium deoxycholate-solubilized apoprotein B (apoB) of native human LDL with preformed, 200 A in diameter, microemulsions of cholesteryl oleate (CO), surface-stabilized by either egg yolk phosphatidylcholine ( EYPC ) or dimyristoyl phosphatidylcholine (DMPC). Gel chromatography of PC/CO/apoB complexes shows co-elution of the complex at 43% PC, 43% CO, and 14% apoB. Negative stain electron microscopy shows the particles to be circular, homogeneous, and approximately 200 A in diameter. PC/CO/apoB complexes exhibit beta-migration on agarose gels and show one high molecular weight protein band on 3.0% sodium dodecyl sulfate-polyacrylamide gels. Differential scanning calorimetry and x-ray scattering show the lipids in the complexes to undergo at least two specific thermal transitions depending on lipid composition, one associated with the core-located cholesterol esters similar to LDL and the protein-free microemulsions and the other from the phospholipid forming the surface monolayer. In addition, particle disruption-protein unfolding/denaturation occur irreversibly at 80-85 degrees C. At 4 degrees C, the secondary structure of apoB on complexes of EYPC /CO/apoB is similar to that of native LDL. For complexes of DMPC/CO/apoB, the secondary structure shows less alpha-helix which correlates with the difference in surface lipid environment. The reassembled complexes of PC/CO/apoB provide a defined system in which the components may be varied systematically in order to study the molecular organization, molecular interactions, and metabolism of LDL.


Comparison of the time course of action of captopril on angiotensin-converting enzyme with the time course of its antihypertensive effect.
The effects of captopril on blood pressure and plasma angiotensin-converting enzyme were studied in cats with two-kidney, one-clip experimental hypertension. The time period for the return of blood pressure to predrug values was observed to be many days longer than the recovery time of enzyme activity. The possibility that the discrepancy might be due to a differential effect of captopril on the pulmonary and vessel wall plasma membrane enzymes as opposed to the circulating plasma enzyme, all of which act on blood-borne angiotensin I, was studied in an anesthetized bioassay cat preparation employing two routes of drug administration. No large disparity between captopril inhibition of the different enzyme systems was noted. It is concluded that the discrepancy between the inhibition of the plasma enzyme and the antihypertensive effect of the drug might result from: (a) a nonspecific return of the disease process that might be predicted to occur with other antihypertensive agents: the "slow pressor" actions of angiotensin are discussed in relationship to this hypothesis; (b) an inhibition of converting enzyme at a localized site, such as the media of the vessel wall or certain brain areas, where the enzyme might not act on blood-borne angiotensin I; and (c) a direct vasodilatory action unrelated to the renin-angiotensin system.


The use of anterior subcutaneous internal fixation (INFIX) for treatment of pelvic ring injuries in major trauma patients, complications and outcomes.
Pelvic anterior internal fixators (INFIX) are a relatively new alternative in the treatment of unstable pelvic fractures. The authors wanted to review the use of complications and outcomes of this method of pelvic fixation at our institution. Patients over the age of 18 who had an INFIX used in treatment of their pelvic ring injury were identified. Patient demographics, fracture type, injury severity score, morbidity, complications and time until removal were recorded. All available patients were followed up following the removal of the INFIX and completed an Iowa Pelvic Score (IPS) at this time. 24 patients (19 male) with a mean age of 38.5 (range 18-71) met the inclusion criteria with an average injury severity score of 29.8 (10-66). The most common complication following insertion was a lateral femoral cutaneous nerve (LFCN) injury, which occurred in 11 patients (bilaterally in two), 6 patients (25%) had ongoing numbness 6 months post removal. Two patients had an infection, one of which prompted the removal of the INFIX. One INFIX was removed for implant failure. All other removals were planned electively. Heterotopic ossification was noted to have occurred in five cases. The mean IPS following removal of the INFIX was 79.2 (52-100). INFIX is a safe and successful treatment for unstable pelvic ring injuries. Overall, patients tolerate the INFIX well with good outcome scores. The main concern being the high rate of LFCN injuries, although many resolved after removal of the INFIX.


Reexamination of effects of stimulus rate and number on the middle components of the averaged electroencephalic response.
The middle components of the evoked cortical response (8-50 msec) were examined under improved signal processing conditions. 512 click stimuli were presented at 5 rates ranging from 1-16/sec to 10 normal-hearing subjects. The influence of stimulus numbers of 32, 64, 128 and 512 and stimulus rates of 1, 2, 4, 8 and 16/sec was examined. Identifiable and repeatable responses were found with as few as 128 stimuli. Stimulus rate had little effect on middle component waveform or its identifiability. An enhancement of signal-to-noise ratio through improved filter conditions is suggested as one reason for the ability to identify middle components to fewer stimuli.


Development of a digital toolkit to improve quality of life of patients with systemic lupus erythematosus.
The purpose of this manuscript is to detail development and initial usability testing of an e-toolkit designed to provide skills and knowledge around self-management behaviors for individuals living with systemic lupus erythematosus. Researchers worked with a steering committee of patients and providers to (1) develop a clickable prototype of an e-toolkit and (2) conduct alpha (individuals not affiliated with an academic clinic as patient or provider) and beta (individual patients with systemic lupus erythematosus as well as members of the clinic healthcare team and individuals who work in patient advocacy organizations) usability testing through semistructured interviews. During the review of the e-toolkit, the feedback provided by participants in both alpha and beta groups centered on two overarching themes: (1) improving user interface and materials and (2) integration of information and supports between toolkit and clinical personnel. Digital approaches that are tailored to individual symptom variation and integrated with a clinical system have the opportunity to enhance ongoing clinical care. These findings support movement toward integrated, team-based care models, tailored digital resources, and use of expanded virtual interaction options to ensure on-going engagement between healthcare providers and systemic lupus erythematosus patients.


Predicting place of death in the program of all-inclusive care for the elderly (PACE): participant versus program characteristics.
To assess the relationship between participant-specific and program-specific characteristics and the place of death in the Program of All-Inclusive Care for the Elderly (PACE). A retrospective, population-based, cross-sectional study. PACE is a community-based managed care model serving a frail, nursing home-eligible population of Medicare beneficiaries. Two thousand two hundred sixty-three PACE decedents from 12 sites that were fully capitated for Medicare and Medicaid by mid-1997. Participant-specific characteristics include the range of demographic, functional, and cognitive status; skilled care needs; medical conditions; and advance care directives measures. The PACE site-specific indicator was also included to assess the effect of program sites on place of death. Bivariate, logistic, and multinomial logit models were employed. The probability of death at home is twice as great (45%) for PACE program participants as for the general population of older Americans. Twenty-one percent of PACE participants die in hospitals, compared with 53% of Medicare beneficiaries. Participants who are older and live in the community have a significantly greater (P< .05) probability of dying at home than does an average PACE enrollee. Those with a do-not-resuscitate order are 7.4% less likely to die in the hospital than are those without. PACE participants with a live-in informal caregiver are 10.3% (P< .05) less likely to die at home than those without a caregiver. When all participant characteristics were accounted for, there remained significant variation in the place of death by PACE plan. We found the least amount of site-specific variation in deaths occurring at home and the most variation in deaths occurring in hospitals. Nationally, the low rate of home death has prompted some to conclude that it is unlikely that we will, as a society, decide to take our dying older people back into our homes. The PACE experience suggests that it is possible to modify this trend. The results indicate that patient and program characteristics are both important in predicting the place of death. The variation across PACE sites suggests that an opportunity for narrowing these gaps, and reducing the proportion of hospital deaths, exists. The PACE end-of-life practice style could potentially be applied to other populations and may have important public policy implications.


Density functional theory study of the interaction of vinyl radical, ethyne, and ethene with benzene, aimed to define an affordable computational level to investigate stability trends in large van der Waals complexes.
Our purpose is to identify a computational level sufficiently dependable and affordable to assess trends in the interaction of a variety of radical or closed shell unsaturated hydro-carbons A adsorbed on soot platelet models B. These systems, of environmental interest, would unavoidably have rather large sizes, thus prompting to explore in this paper the performances of relatively low-level computational methods and compare them with higher-level reference results. To this end, the interaction of three complexes between non-polar species, vinyl radical, ethyne, or ethene (A) with benzene (B) is studied, since these species, involved themselves in growth processes of polycyclic aromatic hydrocarbons (PAHs) and soot particles, are small enough to allow high-level reference calculations of the interaction energy ΔEAB. Counterpoise-corrected interaction energies ΔEAB are used at all stages. (1) Density Functional Theory (DFT) unconstrained optimizations of the A-B complexes are carried out, using the B3LYP-D, ωB97X-D, and M06-2X functionals, with six basis sets: 6-31G(d), 6-311 (2d,p), and 6-311++G(3df,3pd); aug-cc-pVDZ and aug-cc-pVTZ; N07T. (2) Then, unconstrained optimizations by Møller-Plesset second order Perturbation Theory (MP2), with each basis set, allow subsequent single point Coupled Cluster Singles Doubles and perturbative estimate of the Triples energy computations with the same basis sets [CCSD(T)//MP2]. (3) Based on an additivity assumption of (i) the estimated MP2 energy at the complete basis set limit [EMP2/CBS] and (ii) the higher-order correlation energy effects in passing from MP2 to CCSD(T) at the aug-cc-pVTZ basis set, ΔECC-MP, a CCSD(T)/CBS estimate is obtained and taken as a computational energy reference. At DFT, variations in ΔEAB with basis set are not large for the title molecules, and the three functionals perform rather satisfactorily even with rather small basis sets [6-31G(d) and N07T], exhibiting deviation from the computational reference of less than 1 kcal mol(-1). The zero-point vibrational energy corrected estimates Δ(EAB+ZPE), obtained with the three functionals and the 6-31G(d) and N07T basis sets, are compared with experimental D0 measures, when available. In particular, this comparison is finally extended to the naphthalene and coronene dimers and to three π-π associations of different PAHs (R, made by 10, 16, or 24 C atoms) and P (80 C atoms).


Cardiomyocyte Stim1 Deficiency Exacerbates Doxorubicin Cardiotoxicity by Magnification of Endoplasmic Reticulum Stress.
Doxorubicin (Dox) is an effective anticancer agent; however, its cardiotoxicity remains a challenge. Dysfunction of intracellular calcium ion (Ca2+) is implicated in the process of Dox-induced cardiomyocyte apoptosis. Although store-operated Ca2+ entry (SOCE) is suggested to be responsible for Ca2+ entry in cardiomyocytes, the direct role of store-operated Ca2+ channels in Dox-related cardiomyocyte apoptosis is unknown. Cardiomyocyte Stim1-specific knockout or overexpression mice were treated with Dox. Cardiomyocytes were pretreated with Stim1 adenovirus or siRNA followed by Dox incubation in vitro. Cardiac function and underlying mechanisms echocardiography were assessed via immunofluorescence, flow cytometry, real-time PCR, Western blotting and immunoprecipitation. We observed the inhibition of Stim1 expression, association of Stim1 to Orai1 or Trpc1, and SOCE in Dox-treated mouse myocardium and cardiomyocytes. Orai1 and Trpc1 expression remained unchanged. Cardiomyocyte-specific deficiency of Stim1 exacerbated Dox-induced cardiac dysfunction and myocardial apoptosis. However, specific overexpression of Stim1 in the myocardium was associated with amelioration of cardiac dysfunction and myocardial apoptosis. In vitro, STIM1 knockdown potentiated Dox-induced AC16 human cardiomyocyte apoptosis. This apoptosis was attenuated by STIM1 upregulation. Moreover, STIM1 downregulation enhanced Dox-induced endoplasmic reticulum (ER) stress in cardiomyocytes. In contrast, STIM1 overexpression inhibited the activation of the above molecular markers of ER stress. Immunoprecipitation assay showed that STIM1 interacted with GRP78 in cardiomyocytes. This interaction was attenuated in response to Dox treatment. Our data demonstrate that cardiomyocyte STIM1 binding to GRP78 ameliorates Dox cardiotoxicity by inhibiting pro-apoptotic ER stress.


Human eosinophil lysophospholipase: the sole protein component of Charcot-Leyden crystals.
Charcot-Leyden crystals (CLC), formed in vitro from human eosinophils, were recently shown to contain a protein identical in physicochemical characteristics to human eosinophil lysophospholipase. Monospecific antisera, prepared against homogeneous, chromatographically purified eosinophil lysophospholipase and against CLC formed in vitro, yielded precipitin lines fusing in a pattern of immunochemical identity on Ouchterlony analysis with disrupted eosinophils, purified lysophospholipase, and solubilized CLC protein. With antisera to the purified lysophospholipase, CLC present in vivo in human feces were demonstrated by indirect immunofluorescence to contain eosinophil lysophospholipase. Fecal CLC, purified by sequential gradient centrifugation, contained a single protein migrating identically to eosinophil lysophospholipase on SDS polyacrylamide gel electrophoresis. Solubilized fecal CLC were recrystallized to form characteristically-shaped CLC. Thus, naturally occurring CLC are formed solely of human eosinophil lysophospholipase.


A cisplatin-resistant murine leukemia cell line exhibits increased topoisomerase II activity.
cis-Dichlorodiammineplatinum(II) (CDDP) resistance in L1210/10 murine leukemia cells is multifactorial and involves decreased drug uptake, increased glutathione content, and enhanced DNA repair activity. We show here that 0.35 M NaCl nuclear extracts from L1210/10 cells possess an approximately 3-fold increase in DNA topoisomerase II activity, compared with parental L1210 cells, as measured by decatenation of kinetoplast DNA. No difference in topoisomerase I activity is observed between the two cell lines. Immunoblot analysis of topoisomerase II protein in resistant and sensitive cells suggests that the observed differences in topoisomerase II activity cannot be explained by differences in the level of protein expressed. L1210/10 cells are 2.5-fold more sensitive than L1210 cells to the cytotoxic effects of the topoisomerase II inhibitor 4'-(9-acridylamino)methane-sulfon-m-anisidide. Sequential treatment with 4'-(9-acridyl-amino)methanesulfon-m-anisidide and CDDP leads to an additive cytotoxic effect of the two drugs in sensitive L1210 cells, as determined by colony formation in semi-solid medium. In contrast, the same treatment leads to a supra-additive effect in L1210/10 cells, which strongly suggests a role for topoisomerase II in the CDDP resistance of this cell line.


Serum components stimulate pericardial tissue contraction.
Previous experiments have demonstrated the retraction and fibrosis of vital autologous pericardial flap implants in the descending aorta of sheep. An in-vitro model of pericardial tissue contraction was developed that showed healing reactions similar to those observed in the fresh in-vivo flap. Here, the component(s) of serum that stimulate tissue contraction were partially characterized. The molecular weight range and stability (heat and protease resistance) of the serum component(s) are described. Tissue contraction also requires de-novo protein synthesis. Sections (1 cm2) of sheep pericardium were incubated with fractionated, heat-treated, or protease-treated fetal bovine serum for 14 days. In addition, SDS-PAGE protein profiles were generated using tissues incubated with and without cycloheximide for up to 12 days. Tissue contraction was observed in molecular weight serum fractions > or =5 kDa, as well as in samples incubated with heat and protease-treated serum. SDS-PAGE showed the appearance of a protein band after day 4 during the process of tissue contraction that was absent in samples incubated with cycloheximide. Serum fractions > or =5 kDa stimulated protein synthesis and pericardial tissue contraction. The active component(s) was shown to be heat stable, but partially sensitive to protease. The addition of cycloheximide to the culture medium, shown previously to prevent pericardial tissue contraction, inhibited de-novo synthesis of the protein that appeared during the process of tissue contraction.


Glyceraldehyde-3-phosphate dehydrogenase aggregation inhibitor peptide: A potential therapeutic strategy against oxidative stress-induced cell death.
The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has multiple functions, including mediating oxidative stress-induced neuronal cell death. This process is associated with disulfide-bonded GAPDH aggregation. Some reports suggest a link between GAPDH and the pathogenesis of several oxidative stress-related diseases. However, the pathological significance of GAPDH aggregation in disease pathogenesis remains unclear due to the lack of an effective GAPDH aggregation inhibitor. In this study, we identified a GAPDH aggregation inhibitor (GAI) peptide and evaluated its biological profile. The decapeptide GAI specifically inhibited GAPDH aggregation in a concentration-dependent manner. Additionally, the GAI peptide did not affect GAPDH glycolytic activity or cell viability. The GAI peptide also exerted a protective effect against oxidative stress-induced cell death in SH-SY5Y cells. This peptide could potentially serve as a tool to investigate GAPDH aggregation-related neurodegenerative and neuropsychiatric disorders and as a possible therapy for diseases associated with oxidative stress-induced cell death.