Inferior epigastric artery as a conduit for myocardial revascularization.
In 18 patients the inferior epigastric artery (IEA) was used for myocardial revascularization along with the left internal mammary artery and saphenous vein (4.5 grafts/patient). Preoperative duplex scan assessment of IEA correctly predicted its distal diameter. At operation the mean internal diameter was 1.6 (1.5-2.0) mm distally and 2.1 (2.0-2.5) mm proximally. The mean in situ flow after intraluminal injection of papaverine was 115 (36-210) ml/min in IEA and 136 (56-210) ml/min in internal mammary artery. There was no perioperative death or myocardial infarction. Morbidity associated with IEA harvesting was minor: ultrasonography showed diastasis and slower contraction of the rectus muscle at the harvesting site than contralaterally in one case. The preoperative duplex scan was useful for evaluating IEA size. All the patients were clinically improved. The patency rate at selective IEA angiography 3 months post-operatively was 72% (13/18). This poor early patency of IEA in our small series raises doubts concerning the vessel's suitability for myocardial revascularization.


Phase II trial of the pegylated liposomal doxorubicin in previously treated metastatic endometrial cancer: a Gynecologic Oncology Group study.
To determine whether pegylated liposomal doxorubicin (PLD) has antitumor activity in pretreated patients with persistent or recurrent endometrial carcinoma and to define the nature and degree of toxicity of PLD. Women with histologically documented recurrent or persistent measurable endometrial carcinoma and with failure of one prior treatment regardless of prior anthracycline therapy were enrolled. PLD was administered intravenously over a 1-hour period at a dose of 50 mg/m(2) every 4 weeks; the dosage was modified in accordance with observed toxicity. Of 46 patients entered, 42 were assessable for response, as three were declared ineligible on central pathology review and one was not assessable for response. Forty had received prior chemotherapy, 11 hormonal therapy, and 29 radiation therapy. Doxorubicin had been given to 32 patients, carboplatin with paclitaxel to six, carboplatin to one, and fluorouracil to one. Four patients had partial responses lasting 1.1, 2.1, 3.3, and 5.4 months; the overall response rate was 9.5% (95% confidence interval, 2.7% to 22.6%). Three of these responses (in liver and in lymph node) occurred in patients who had progressed after doxorubicin with either paclitaxel or cisplatin. The median number of courses was 2.5 (range, one to 14). Toxicity was generally mild: only 25 patients experienced leukopenia, with a median WBC count of 2,900 (range, 800 to 3,900) at nadir. The only grade 4 toxicities were one episode each of esophagitis, hematuria, and vomiting. The median overall survival was 8.2 months. PLD has only limited activity in pretreated advanced, recurrent endometrial cancer, but further trials in anthracycline-naive patients and in previously untreated patients are ongoing. Its toxicity profile should permit its use in combination with myelosuppressive drugs.


Chromosomal mosaicism in amniotic cell culture. A diagnostic Dilemma.
Chromosomal mosaicism, with supernumerary C or C group elements, was observed in the cultured amniotic fluid cells of four fetuses undergoing prenatal diagnosis. There were discrepancies in the karyotypes of cells from different culture flasts of a single fluid sample, between cultures from successive uterine aspirations, and between cultures of amniotic fluid and fetal tissues obtained by pregnancy interruption. Most alarming was the finding, in one case, of identical mosaicism in different culture flasks as well as in repeated amniocenteses derived from a phenytypically normal fetus. In each instance, the aborted fetus or newborn infant proved to be phenotypically and cytogenetically normal. This suggests that mosaicism, demonstrated in cultured amniotic fluid cells, may not reflect the chromosomal constitution of the fetus and may pose a dilemma regarding an accurate diagnosis.


SQUIRE 2.0 (Standards for QUality Improvement Reporting Excellence): revised publication guidelines from a detailed consensus process.
In the past several years, the science of health care improvement has advanced considerably. In this article, we describe the development of SQUIRE 2.0 and its key components. We undertook the revision between 2012 and 2015 using (1) interviews and focus groups to evaluate SQUIRE 1.0 plus feedback from an international steering group, (2) face-to-face consensus meetings to develop interim drafts, and (3) pilot testing with authors and a public comment period. SQUIRE 2.0 emphasizes 3 key components of systematic efforts to improve the quality, value, and safety of health care: formal and informal theory in planning, implementing, and evaluating improvement work; the context in which the work is done; and the study of the intervention(s). SQUIRE 2.0 is intended for reporting the range of methods used to improve health care, recognizing that they can be complex and multidimensional. It provides common ground to share these discoveries in the scholarly literature (www.squire-statement.org).


When general practitioners don't feel appreciated by their patients: prospective effects on well-being and work-family conflict in a Swiss longitudinal study.
Impaired well-being and high work-family conflict are critical issues among GPs. This research examined an understudied psychosocial risk factor for these outcomes, namely GPs' perception that they invest more in the relationship with their patients than what they receive in return (i.e. lack of reward in their relationship with patients). To test the effect of lack of reward as a risk factor for poor well-being and work-family conflict among GPs. Longitudinal study (12 months time lag). 272 GPs in Switzerland [mean age 54.5 (SD = 8.3), 73% male] volunteered to participate in the study. 270 participants completed the baseline survey and 252 completed the follow-up survey. Of these, six retired between the baseline and the follow-up survey, resulting in a sample size of 246 participants at t2. Outcome measures were burnout, sleep problems, self-perceived health and work-family conflict. Strength and direction of prospective effects were tested using cross-lagged models. Lack of reward was related to an increase in emotional exhaustion (β = 0.15), sleep problems (β = 0.16) and work-family conflict (β = 0.19) and a decrease in self-perceived health (β = -0.17). Effects on depersonalization and personal accomplishment were not significant. Regarding reversed effects of impaired well-being on lack of reward, emotional exhaustion (β = 0.14) and self-perceived health (β = -0.13) predicted future level of lack of reward. Lack of reward by patients is a risk factor in GPs' mental health.


Synchronization of the circadian clock to the environment tracked in real time.
The circadian system of the cyanobacterium Synechococcus elongatus PCC 7942 relies on a three-protein nanomachine (KaiA, KaiB, and KaiC) that undergoes an oscillatory phosphorylation cycle with a period of ~24 h. This core oscillator can be reconstituted in vitro and is used to study the molecular mechanisms of circadian timekeeping and entrainment. Previous studies showed that two key metabolic changes that occur in cells during the transition into darkness, changes in the ATP/ADP ratio and redox status of the quinone pool, are cues that entrain the circadian clock. By changing the ATP/ADP ratio or adding oxidized quinone, one can shift the phase of the phosphorylation cycle of the core oscillator in vitro. However, the in vitro oscillator cannot explain gene expression patterns because the simple mixture lacks the output components that connect the clock to genes. Recently, a high-throughput in vitro system termed the in vitro clock (IVC) that contains both the core oscillator and the output components was developed. Here, we used IVC reactions and performed massively parallel experiments to study entrainment, the synchronization of the clock with the environment, in the presence of output components. Our results indicate that the IVC better explains the in vivo clock-resetting phenotypes of wild-type and mutant strains and that the output components are deeply engaged with the core oscillator, affecting the way input signals entrain the core pacemaker. These findings blur the line between input and output pathways and support our previous demonstration that key output components are fundamental parts of the clock.


Molecular cloning of a xylanase gene from Bacteroides succinogenes and its expression in Escherichia coli.
A gene coding for xylanase synthesis in Bacteroides succinogenes was isolated by cloning, with Escherichia coli HB101 as the host. After partial digestion of B. succinogenes DNA with Sau3A, fragments were ligated into the BamHI site of pBR322 and transformed into E. coli HB101. Of 14,000 colonies screened, 4 produced clear halos on Remazol brilliant blue-xylan agar. Plasmids from two stable clones recovered exhibited identical restriction enzyme patterns, with the same 9.4-kilobase-pair (kbp) insert. The plasmid was designated pBX1. After subcloning of restriction enzyme fragments, a 3-kbp fragment was found to code for xylanase activity in either orientation when inserted into pUC18 and pUC19. The original clone possessed approximately 10-fold higher xylanase activity than did clones harboring the 3-kbp insert in pUC18, pUC19, or pBR322. The enzyme was partially secreted into the periplasmic space of E. coli. The periplasmic enzyme of the BX1 clone had 2% of the activity on carboxymethyl cellulose and less than 0.2% of the activity on p-nitrophenyl xyloside and a range of other substrates that it exhibited on xylan. The xylanase gene was not subject to catabolite repression by glucose or induction by either xylan or xylose. The xylanase activity migrated as a single broad band on nondenaturing polyacrylamide gels. The Km of the pBX1-encoded enzyme was 0.22% (wt/vol) of xylan, which was similar to that for the xylanase activity in an extracellular enzyme preparation from B. succinogenes. Based on these data it appears that the xylanase gene expressed in E. coli is fully functional and codes for an enzyme with properties similar to the B. succinogenes enzyme(s).


Discovery of a Potent and Selective Steroidal Glucocorticoid Receptor Antagonist (ORIC-101).
The glucocorticoid receptor (GR) has been linked to therapy resistance across a wide range of cancer types. Preclinical data suggest that antagonists of this nuclear receptor may enhance the activity of anticancer therapy. The first-generation GR antagonist mifepristone is currently undergoing clinical evaluation in various oncology settings. Structure-based modification of mifepristone led to the discovery of ORIC-101 (28), a highly potent steroidal GR antagonist with reduced androgen receptor (AR) agonistic activity amenable for dosing in androgen receptor positive tumors and with improved CYP2C8 and CYP2C9 inhibition profile to minimize drug-drug interaction potential. Unlike mifepristone, 28 could be codosed with chemotherapeutic agents readily metabolized by CYP2C8 such as paclitaxel. Furthermore, 28 demonstrated in vivo antitumor activity by enhancing response to chemotherapy in the GR+ OVCAR5 ovarian cancer xenograft model. Clinical evaluation of safety and therapeutic potential of 28 is underway.


Numerical taxonomy of gram-positive and catalase-positive rods isolated from frozen vegetables.
One hundred isolates from peas, beans, and corn were compared with cultures of Corynebacterium, Microbacterium, and Arthrobacter by use of numerical taxonomic procedures. Six groups (clusters), representing 75% of the isolates, resembled Corynebacteriaceae. There was some doubt regarding the genera represented because the groups were not closely related to many of the known strains. The relationship of the different groups to each other, as well as a number of their properties, is presented.


Lorvotuzumab mertansine: antibody-drug-conjugate for CD56+ multiple myeloma.
Lorvotuzumab mertansine (LM) is an ADC composed of an anti CD56 humanized N901 monoclonal antibody conjugated via a stable disulfide linker to the maytansinoid DM1. CD56 is expressed in up to 78% of multiple myelomas. LM displays antitumor activity in preclinical models of multiple myeloma. In a phase I study of MM, the MTD of single-agent LM was 112 mg/m². The dose-limiting toxicities were grade 3 fatigue and grade 3 acute, reversible, renal failure. 2 PRs and 4 MRs were observed at various dose levels starting at 60 mg/m². Building on the single agent experience, a phase II study of LM in combination with lenalidomide and dexamethasone was conducted. The optimal dose of LM was 75 mg/m² in the combination. The ORR was 56.4%. The most common treatment-related AE was peripheral neuropathy (PN), mostly grade 2 or less, with the majority of patients having a grade 1 PN at baseline. Continued evaluation of optimal dosing levels and schedules will be important to better define the utility of this promising treatment.


bcl-2 overexpression promotes myocyte proliferation.
To determine the influence of Bcl-2 on the developmental biology of myocytes, we analyzed the population dynamics of this cell type in the heart of transgenic (TG) mice overexpressing Bcl-2 under the control of the alpha-myosin heavy chain promoter. TG mice and non-TG (wild type, WT) mice were studied at 24 days, 2 months, and 4 months after birth. Bcl-2 overexpression produced a significant increase in the percentage of cycling myocytes and their mitotic index. These effects were strictly connected to the expression of the transgene, as demonstrated in isolated myocytes. The formation of mitotic spindle and contractile ring was identified in replicating cells. These typical aspects of mitosis were complemented with the demonstration of karyokinesis and cytokinesis to provide structural evidence of cell division. Apoptosis was low at all ages and was not affected by Bcl-2. The higher cell replication rate in TG was conditioned by a decrease in the expression of the cell-cycle inhibitors, p21(WAF1) and p16(INK4a), and by an increase in Mdm2-p53 complexes. In comparison with WT, TG had 0.4 x 10(6), 0.74 x 10(6), and 1.2 x 10(6) more myocytes in the left ventricle at 24 days, 2 months, and 4 months, respectively. Binucleated myocytes were 12% and 25% larger in WT than in TG mice at 2 and 4 months of age. Taken together, these observations reveal a previously uncharacterized replication-enhancing function of Bcl-2 in myocytes in vivo in the absence of stressful conditions.


High-dose-level effects in mutagenicity assays utilizing mammalian cells in culture.
We report here results obtained using sodium chloride and potassium chloride as model compounds to investigate the effects of high dose levels and osmolarity-mediated artefacts in mutagenicity assays using cultured mammalian cells. Three assay systems were used with different genetic end-points: (i) mutation to 6-thioguanine resistance in Chinese hamster V79 cells; (ii) induction of chromosome aberrations in Chinese hamster ovary cells; and (iii) induction of unscheduled DNA synthesis in HeLa S3 cells. In V79 cells we observed sporadic increases in mutation frequency after treatment with sodium chloride. Potassium chloride increased the mutation frequency in a narrow interval of dose levels. Chromosome aberrations were induced by both compounds at the highest concentrations tested, confirming previously published data; the effects of potassium chloride were more marked. The chromosome aberrations observed included both deletions and exchange figures. Neither compound induced UDS in our experiments. We performed the Ames test as a check for mutagenic contaminants and both compounds gave entirely negative results. Although the osmolarities of sodium chloride and potassium chloride solutions were similar, the effects of potassium chloride were always greater at equivalent concentrations; the nature of the solute, and not only the observed osmolarity, appears to influence the results obtained. Similarly, we have observed that dimethylsulphoxide causes a marked increase in the osmolarity of the culture medium without any obvious induction of chromosome aberrations. Although the presence of S9 had little effect on the osmolarity of the medium, differences in response were observed after treatments with or without S9. For the two model compounds it appears that an osmotic mechanism alone does not provide a sufficient explanation of the findings.


[Spectrum-activity relationship of trichosanthis fructus and trichosanthis fructus strip pieces for rat myocardial ischemia-reperfusion injury].
To investigate the spectrum-activity relationship of Trichosanthis Fructus and Trichosanthis Fructus strip pieces for rat myocardial ischemia-reperfusion injury. HPLC fingerprints of Trichosanthis Fructus and Trichosanthis Fructus strip pieces were established, and the values of creatinekinase-MB (CK-MB), myoglobin (MYO) and cardiac troponin-T (cTNT) in 3 dose groups (2.25, 13.5, 27.0 g·kg⁻¹, equivalent to the crude herb g·kg⁻¹) of Trichosanthis Fructus and Trichosanthis Fructus strip pieces with myocardial ischemia-reperfusion injury in rats were measured, and the grey relational analysis was used to study the spectrum-activity relationship of Trichosanthis Fructus and Trichosanthis Fructus strip pieces for rat myocardial ischemia-reperfusion injury. With the dosage increase from 2.25 g·kg⁻¹ to 27.0 g·kg⁻¹, the correlation degree of spectrum-activity relationship of Trichosanthis Fructus and Trichosanthis Fructus strip pieces was also enhanced, but the change trend was different between these two groups. According to the frequency of the top 10 peaks in the correlation degree, peak 17, 14, 16, 19, 32, 12, 26, 30, 4, 6 and 2 were the basic effective substances group of Trichosanthis Fructus, peak 6,14,12,32,30,4 and 6 were the basic effective substances group of Trichosanthis Fructus strip pieces. Peak 6, 14, 12, 32, 30, 4 and 26 in fingerprints of Trichosanthis Fructus and Trichosanthis Fructus strip pieces were the main common pharmacodynamic substance base, among them, peak 6 was 5-hydroxymethyl furfural, peak 14 was vanillic acid and the peak 28 was rutin, but the correlation degree with the efficacy was different. The effect of Trichosanthis Fructus and Trichosanthis Fructus strip pieces on rat myocardial ischemia-reperfusion injury was due to the synergistic effect of the effective substance groups related to the dosage. The essential pharmacodynamic substance groups of Trichosanthis Fructus and Trichosanthis Fructus strip pieces were different, but they shared a common active ingredient group.


[Construction and expression of six deletion mutants of human astrovirus C-terminal nsP1a/4 protein].
Human astrovirus (HAstV) is one of the leading causes of actue virual diarrhea in infants. HAstV-induced epithdlial cell apoptosis plays an important role in the pathogenesis of HAstV infection. Our previous study indicated that HAstV non-structural protein nsPla C-terminal protein nsPla/4 was the major apoptosis functional protein and probably contained the main apoptosis domains. In order to screen for astrovirus encoded apoptotic protien, nsPla/4 and six turncated proteins, which possessed nsPla/4 protein different function domain ,were cloned into green fluorescent protein (GFP) vector pEG-FP-N3. After 24-72 h transfection, the fusion protein expression in BHK21 cells, was analysis by fluorescence microscope and Western blot. The results indicated seven fusion proteins were observed successfully in BHK21 cell after transfected for 24 h. Western blot analysis showed that the level of fusion protein expressed in BHK21 cells was increased significantly at 72h compared to 48h in transfected cells. The successful expression of deletion mutants of nsPla/4 protein was an important foundation to gain further insights into the function of apoptosis domains of nsPla/4 protein and it would also provide research platform to further confirm the molecule pathogenic mechanism of human astrovirus.


Pharmacogenomics of multifactorial diseases: a focus on psoriatic arthritis.
This review will outline the current pharmacogenomics knowledge about psoriatic arthritis with a special attention to the perspectives and the challenges for its implementation in the clinical practice. To date, different drugs have been developed to contrast the symptoms and the progression of psoriatic arthritis. However, patients have shown high variability of drug response in relation to their genetic makeup. In this context, the advances made in the knowledge and the potentialities of genome-drugs associations paved the path for the development of a precision medicine. In fact, these associations may be successfully combined with the environment information to provide new strategies able to prevent and improve the disease management as well as to enhance the patients quality of life.


The Relationship between Objectively Measured and Self-Reported Sedentary Behaviours and Social Connectedness among Adolescents.
Adolescents spend significant amounts of time engaged in various types of sedentary behaviour (SB). This study examined associations between adolescents' objectively measured sedentary time, sitting time, specific self-reported SBs and social connectedness. Adolescents (n = 429, 15.5 years, 41% male) completed an online survey reporting time in seven SBs (TV/videos/DVDs, computer/video games, internet, homework, reading, car and bus travel; examined individually and summed for screen time and total SB), and social connectedness using the eight-item Social Connectedness Scale. A subsample (n = 353) also wore an ActiGraph GT3X+ (model GT3X+, Pensacola, FL, USA) accelerometer to measure sedentary time (<100 cpm) and n = 237 wore an activPAL (PAL Technologies Ltd., Glasgow, Scotland) inclinometer to measure sitting time. Multiple linear mixed models determined associations between each SB variable and social connectedness, adjusting for confounders. Adolescents spent on average 7.8 h/day in self-reported total SB, 4.4 h/day in screen time, 9.1 h/day in ActiGraph-measured sedentary time, and 9.5 h/day in activPAL-measured sitting time. After adjusting for age, sex and area level socioeconomic status, total SB (-0.24, 95%CI: -0.37, -0.11), screen time (-0.23, 95%CI: -0.41, -0.05) and two individual SBs (computer/video games (-1.07, 95%CI: -1.53, -0.60), homework (-0.61, 95%CI: ⁻1.04, -0.18) were negatively associated with social connectedness. There were no associations with the objective measures. The relationships may be bi-directional; therefore, future research should involve longitudinal designs and explore other potential contributing factors.


Problems involved with the use of comforters.
Comforters have been associated with three areas of concern for the dental profession: rampant caries, malocclusion and gingival recession. The most serious of these, rampant caries, is associated with the practice of sweetening the comforter. Every effort should be made to eliminate this hazard to the dentition by means of dental health education for the community, especially of pregnant and nursing mothers. An open question remains on whether unsweetened dummy sucking should be promoted in order to intercept malocclusions likely to arise from digit sucking.


Preparation of active enzyme samples for IR studies of Na+/K+-ATPase.
In the case of the integral membrane protein Na+/K+-ATPase, preparation of highly concentrated samples for IR difference spectroscopy often leads to inactivation of the enzyme. Therefore, we compared the activity of Na+/K+-ATPase using different techniques of sample preparation. The loss of activity can be minimized by cooling the sample to 10 degrees C and by the addition of glycerol and dithiothreitol. The activity of Na+/K+-ATPase isolated from pig kidney is independent of the protein concentration whereas the enzyme from shark rectal gland is inactivated at concentrations above 1 microg/microL and is thus unsuitable for IR experiments.


Extramural US Federal Research Grants For Health Outcomes Associated With Climate Change Inadequate, Too Narrow In Focus.
Climate change causes and exacerbates disease, creates and worsens health disparities, disrupts health care delivery, and imposes a significant disease burden in the US and globally. Critical knowledge gaps hinder an evidence-based response and are perpetuated by scarce federal research funds. We identified and described extramural US federal research funding (that is, grants provided to organizations and institutions outside of federal agencies) that both addressed health outcomes associated with climate change and was awarded between 2010 and 2020. During this eleven-year period, 102 grants met our criteria, totaling approximately $58.7 million, or approximately $5.3 million per year (2020 adjusted US dollars). Federal investments in climate change and health research during this period failed to address the breadth of climate-sensitive exposures, health outcomes, and impacts on vulnerable populations. Moving forward, in addition to increasing investment in climate and health research across all known hazards, critical attention should be placed on vulnerable populations and health equity. To achieve this, increased federal research coordination and cooperation are needed, as well as a mechanism to track this funding.


Expression and localization of gap junctional connexins 26 and 43 in bovine periovulatory follicles and in corpus luteum during different functional stages of oestrous cycle and pregnancy.
The aim of this study was to characterize the regulation of connexins (Cx26 and Cx43) in the bovine ovary (experiment 1-3). Experiment 1: ovaries containing preovulatory follicles or corpora lutea (CL) were collected at 0, 4, 10, 20, 25 (follicles) and 60 h (CL) relative to injection of GnRH. Experiment 2: CL were assigned to the following stages: days 1-2, 3-4, 5-7, 8-12, 13-16, >18 (after regression) of oestrous cycle and of early and late pregnancy (<4 and >4 months). Experiment 3: induced luteolysis, cows on days 8-12 were injected with PGF2alpha analogue (Cloprostenol), and CL were collected by transvaginal ovariectomy before and 0.5, 2, 4, 12, 24, 48 and 64 h after PGF2alpha injection. Real-time RT-PCR was applied to investigate mRNA expression and immunofluorescence was utilized for protein localization. Cx26 mRNA increased rapidly 4 h after GnRH injection (during LH surge) and decreased afterwards during the whole experimental period. Cx43 mRNA expression decreased continuously after GnRH application. Cx26 mRNA in CL increased significantly in the second part of oestrous cycle and after regression. In contrast, the highest mRNA expression for Cx43 in CL was detected during the early luteal phase. After induced luteolysis the mRNA expression of Cx26 increased significantly at 24 h. As shown by immunofluorescence, Cx26 was predominantly localized in the connective tissue and blood vessels of bovine CL, whereas Cx43 was present in the luteal cells and blood vessels. This resulted in a strong increase of Cx26 expression during the late luteal phase and after luteal regression. Subsequently, Cx43 expression was distinctly decreased after luteal regression. These data suggest that Cx26 and Cx43 are involved in the local cellular mechanisms participating in tissue remodelling during the critical time around periovulation as well as during CL formation (angiogenesis), function and regression in the bovine ovary.


Tough trade-offs: medical bills, family finances and access to care.
About 20 million American families-representing 43 million people-reported problems paying medical bills in 2003, according to a new study by the Center for Studying Health System Change (HSC). While uninsured families are more likely to have medical bill problems, two-thirds of families with problems paying medical bills have health insurance coverage. Of all families with medical bill problems, almost two-thirds reported difficulty paying for other basic necessities-rent, mortgage payments, transportation or food-as a result of medical debt. People in families with medical bill problems also reported much greater trouble getting care because of cost concerns-one in three did not get a prescription drug, one in four delayed care and one in eight went without needed care.


Benzene toxicity.
The proper control of benzene in the workplace and the general environment has been a subject of relatively intense interest for at least the past decade. The author reviews the toxicology of benzene, its hematologic effects in the human, and methods of measurement of benzene exposure in the workplace.


Dynamic analysis of genetic control and regulation of amino acid synthesis: the tryptophan operon in Escherichia coli.
A mathematical model of the tryptophan operon is analyzed to investigate the regulatory effects of feedback repression and the demand for tryptophan in the cell. In this model, feedback repression is considered to be a two-step process. First, the endproduct tryptophan combines with the inactive repressor produced by the regulatory genes to yield an active complex. This complex subsequently binds to the operator and prevents transcription of the structural genes into mRNA. The demand for tryptophan in the cell is modeled by a hyperbolic saturation function of the Michaelis-Menten type. Results are obtained for the expression of the tryptophan operon in Escherichia coli and their applicability to tryptophan production by microbial fermentation is discussed. It is shown that, depending on the strain level of the operon and the rate of utilization of tryptophan in the cell, an overproduction of tryptophan can be achieved under stable operating conditions; in other circumstances, the operon may become stable or unstable, and may lead to a periodic synthesis.


Prevalence and correlates of receiving and sharing high-penetrance cancer genetic test results: findings from the Health Information National Trends Survey.
The aim of this study was to explore the prevalence and correlates of receiving and sharing high-penetrance cancer genetic test results. Participants completed the population-based, cross-sectional 2013 Health Information National Trends Survey. We examined sociodemographic characteristics of participants reporting having had BRCA1/2 or Lynch syndrome genetic testing, and sociodemographic and psychosocial correlates of sharing test results with health professionals and family members. Participants who underwent BRCA1/2 or Lynch syndrome genetic testing (n = 77; 2.42% of respondents) were more likely to be female and to have a family or personal history of cancer than those not undergoing testing. Approximately three-quarters of participants shared results with health professionals and three-quarters with their family; only 4% did not share results with anyone. Participants who shared results with health professionals reported greater optimism, self-efficacy for health management, and trust in information from their doctors. Participants who shared results with their family were more likely to be female and to have a personal history of cancer, and had greater self-efficacy for health management, perceived less ambiguity in cancer prevention recommendations, and lower cancer prevention fatalism. We identified several novel psychosocial correlates of sharing genetic information. Health professionals may use this information to identify patients less likely to share information with at-risk family members.


The radioimmunoassay of human urinary kallikrein and comparisons with kallikrein activity measurements.
Human urinary kallikrein was purified to homogeneity, and an antiserum to it was raised in rabbits. A RIA was devised which uses this rabbit antiserum (Keq = 2.75 x 10(11) M-1) in a final dilution of 1:2,500,000 and the purified kallikrein labeled with 125I using a lactoperoxidase method. Assay sensitivity is 8 pg kallikrein. Thus far, the assay is specific for human and perhaps monkey urinary kallikrein. Correlations between this assay of immunoreactive kallikrein and the alpha-N-Tosyl-L-arginine-[3H]methylester (Tos-Arg-OMe) activity method or a kininogenase assay were highly significant (r = 0.94 and 0.96, respectively) and show that each assay measures human urinary kallikrein comparably. Low or high dietary sodium intakes, maneuvers known to change human urinary Tos-Arg-OMe esterase excretion, change immunoreactive kallikrein to an equivalent degree. Normal black children, already known to excrete significantly less Tos-Arg-OMe esterase than white children, excrete similarly reduced amounts of immunoreactive kallikrein. Kallikrein excretion in children with cystic fibrosis of the pancreas was not different from that in normal children. The data show that a specific and sensitive direct RIA for human urinary kallikrein has been developed and that both the Tos-Arg-OMe esterase and kininogenase assays measure human urinary kallikrein activity specifically, at least in the described circumstances.


Optimization of an HPLC method for determination of gabapentin in dosage forms through derivatization with 1-fluoro-2,4-dinitrobenzene.
A rapid, sensitive and accurate high performance liquid chromatography with UV detection method was developed and validated for the quantification of gabapentin in dosage forms. Gabapentin was quantified after pre-column derivatization with 1-fluoro-2,4-dinitrobenzene. Amlodipine was used as an internal standard. The chromatographic separation was carried out on a Nova-Pak C(18) column using a mixture of acetonitrile-sodium dihydrogenphosphate (pH 2.5; 0.05 M) (70:30, v/v) as mobile phase with UV detection at 360 nm. The method was linear over the range of 10-500 microg/ml of gabapentin (r(2)>0.999). The within-day and between-day precision values were in the range of 0.86-1.11%. The method was successfully used for quantitative determination and dissolution rate study of Neurontin capsules.


Patient Access, Unmet Medical Need, Expected Benefits, and Concerns Related to the Utilisation of Biosimilars in Eastern European Countries: A Survey of Experts.
This policy research aims to map patient access barriers to biologic treatments, to explore how increased uptake of biosimilars may lower these hurdles and to identify factors limiting the increased utilisation of biosimilars. A policy survey was developed to review these questions in 10 Central and Eastern European (CEE) and Commonwealth of Independent States (CIS) countries. Two experts (one public and one private sector representative) from each country completed the survey. Questions were related to patient access, purchasing, clinical practice, and real-world data collection on both original biologics and biosimilars. Restrictions on the number of patients that can be treated and related waiting lists were reported as key patient access barriers. According to respondents, for both clinicians and payers the primary benefit of switching patients to biosimilars would be to treat more patients. However, concerns with therapeutic equivalence and fear of immunogenicity may reduce utilisation of biosimilars. Similar limitations in patient access to both original biologics and biosimilars raise concerns about the appropriateness and success of current biosimilar policies in CEE and CIS countries. The conceptual framework for additional real-world data collection exists in all countries which may provide a basis for future risk-management activities including vigorous pharmacovigilance data collection.


Cobalt-Catalyzed Alkylation of Aromatic Amines by Alcohols.
The implementation of inexpensive, Earth-abundant metals in typical noble-metal-mediated chemistry is a major goal in homogeneous catalysis. A sustainable or green reaction that has received a lot of attention in recent years and is preferentially catalyzed by Ir or Ru complexes is the alkylation of amines by alcohols. It is based on the borrowing hydrogen or hydrogen autotransfer concept. Herein, we report on the Co-catalyzed alkylation of aromatic amines by alcohols. The reaction proceeds under mild conditions, and selectively generates monoalkylated amines. The observed selectivity allows the synthesis of unsymmetrically substituted diamines. A novel Co complex stabilized by a PN5 P ligand catalyzes the reactions most efficiently.


Percutaneous interventions.
The catheterization laboratory is no longer limited to coronary arterial interventions. Physicians have been striving to make more interventions less invasive that now can include carotid, renal, and peripheral arterial stenting, as well as less invasive repair of atrial septal abnormalities. Even cardiac assist devices can be implanted percutaneously to bridge a critically ill patient to other modes of treatment. This article will give a brief overview of each intervention and identify important nursing care.


The NH(2)-terminal region of Streptococcus pyogenes M5 protein confers protection against degradation by proteases and enhances mucosal colonization of mice.
The NH(2)-terminal sequence of the M protein from group A streptococci defines the serotype of the organism and contains epitopes that evoke bactericidal antibodies. To identify additional roles for this region of the M protein, we constructed a mutant of M5 group A streptococci expressing an M protein with a deletion of amino acid residues 3-22 (DeltaNH(2)). M5 streptococci and the DeltaNH(2) mutant were resistant to phagocytosis and were similarly virulent in mice. However, DeltaNH(2) was significantly less hydrophobic, contained less lipoteichoic acid on its surface, and demonstrated reduced adherence to epithelial cells. These differences were abolished when organisms were grown in the presence of protease inhibitors. Treatment with cysteine proteases or with human saliva resulted in the release of M protein from the DeltaNH(2) mutant at a significantly greater rate than observed with the wild-type M5 strain. Compared with the parent strain, the DeltaNH(2) strain also showed a significant reduction in its ability to colonize the upper respiratory mucosa of mice. The NH(2) terminus of M5 protein has an important role in protecting the surface protein from proteolytic cleavage, thus preserving its function as an anchor for lipoteichoic acid, which is a primary mediator of adherence to epithelial cells and colonization.


Agent-based Modeling in Tobacco Regulatory Science: Exploring 'What if' in Waterpipe Smoking.
Waterpipe tobacco smoking (WTS) is an emerging public health crisis, particularly among youth and young adults. Different from the use of other tobacco products and e-cigarettes, WTS tends to be a social activity occurring among friends or persons associated with social networks. In this paper, we review a potential strategy for WTS-related research. As a bottom-up computational model, agent-based modeling (ABM) can simulate the actions and interactions of agents, as well as the dynamic interactions between agents and their environments, to gain an understanding of the functioning of a system. ABM is particularly useful for incorporating the influence of social networks in WTS, and capturing people's space-time activity and the spatial distribution of WTS venues. Comprehensive knowledge of WTS-related behaviors at the individual level is needed to take advantage of ABM and use it to examine policies such as the interaction between WTS and cigarette smoking and the effect of flavors used in waterpipe tobacco. Longitudinal and WTS-specific surveys and laboratory experiments are particularly helpful to understand WTS basic mechanisms and elicit individual preferences, respectively. We argue that the uniqueness of WTS makes ABM a promising tool to be used in WTS-related research, as well as understanding use of other tobacco products.


Escherichia coli DnaX product, the tau subunit of DNA polymerase III, is a multifunctional protein with single-stranded DNA-dependent ATPase activity.
The dnaZX gene of Escherichia coli directs the synthesis of two proteins, DnaZ and DnaX. These products are confirmed as the gamma and tau subunits of DNA polymerase III because antibody to a synthetic peptide present in both the DnaZ and DnaX proteins reacts also with the gamma and tau subunits of holoenzyme. To characterize biochemically the tau subunit, for which there has been no activity assay, the dnaZX gene was fused to the beta-galactosidase gene to encode a fusion product in which the 20 C-terminal amino acids of the DnaX protein (tau) were replaced by beta-galactosidase lacking only 7 N-terminal amino acids. The 185-kDa fusion protein, which retained beta-galactosidase activity, was overproduced to the level of about 5% of the soluble cellular protein by placing the gene fusion under control of the tac promoter and Shine-Dalgarno sequence. The fusion protein was isolated in one step by affinity chromatography on p-aminobenzyl 1-thio-beta-D-galactopyranoside-agarose. The purified fusion protein also had ATPase (and dATPase) activity that was dependent on single-stranded DNA. This activity copurified with the beta-galactosidase activity not only through the affinity column but also through a subsequent gel filtration. We conclude that the DnaX protein function involves binding to single-stranded DNA and hydrolysis of ATP or dATP, in addition to binding to other DNA polymerase III holoenzyme components, increasing the processivity of the core enzyme, and serving as a substrate for the production of the gamma subunit.


Stridor: an unexpected complication from chemoradiotherapy for anal cancer.
The treatment of choice for anal cancer is chemoradiotherapy. Skin reaction and bowel symptoms such as tenesmus, diarrhoea and bleeding are common side effects. We report a patient who developed stridor as a result of chemoradiotherapy for anal cancer and discuss the pathogenesis and potential consequences.


[Overexpression of human EP4 receptor in vascular smooth muscle cells attenuates angiotensin II-induced hypertension in mice].
Prostaglandin E2 (PGE2) plays an important role in cardiovascular system. PGE2 regulates blood pressure through its 4 G protein coupled receptors, i.e., EP1, EP2, EP3, and EP4. The aim of this study was to investigate the role of EP4 receptors in vascular smooth muscle cells (VSMC) in blood pressure regulation. VSMC-specific human EP4 transgenic (VSMC-hEP4 Tg) mice were generated and genotyped. The systolic blood pressure (SBP) of the VSMC-hEP4 Tg mice and the wild-type (WT) littermates was measured under normal, low-salt (LSD) and high-salt diet (HSD) conditions using a tail-cuff method. Both WT and VSMC-hEP4 Tg mice were administered with a chronic infusion of angiotensin II (Ang II) with an osmotic pump and SBP levels were monitored every week. The mean arterial blood pressure (MAP) of WT and VSMC-hEP4 Tg mice upon Ang II intravenous infusion was measured via carotid arterial catheterization. Ang II-induced vasoconstriction of the mesenteric arterial rings from WT and VSMC-hEP4 Tg mice was measured using the multi myograph system. The effect of PGE1-OH (a selective EP4 agonist) on Ang II-induced phosphorylation of myosin phosphatase target subunit 1 (MYPT1) was detected by Western blot. The effect of two additional EP4 specific agonists (CAY10580 and CAY10598, 0.5 mg/kg) on blood pressure of WT mice was measured by carotid arterial catheterization. The results showed that the VSMC-hEP4 Tg mice were successfully generated and their basal SBP levels were lower than those of WT mice. Although blood pressure levels were significantly altered in WT mice under LSD and HSD, little change was observed in the VSMC-hEP4 Tg mice. After a chronic infusion and an acute intravenous injection of Ang II, SBP levels of VSMC-hEP4 Tg mice were significantly lower than those of WT mice. In addition, both CAY10580 and CAY10598 significantly reduced MAP levels of WT mice. Ex vivo study showed that treatment of isolated mesenteric arteries with PGE1-OH inhibited Ang II-induced phosphorylation of MYPT1. Collectively, these results demonstrate that specific overexpression of human EP4 gene in VSMCs significantly reduces basal blood pressure levels and attenuates Ang II-induced hypertension, possibly via inhibiting Ang II/AT1 signaling pathway. Our findings suggest that EP4 may represent an attractive target for the treatment of hypertension.


FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.
AHSG, a serum glycoprotein with recognized anti-calcification activity, has also been suggested to modulate both bone formation and resorption. Though the bulk of AHSG is mostly synthesized in the liver, it has been claimed that also bone cells might produce it. However, the extent of the bone AHSG production and the potential controlling factors remain to be definitively proven. A relevant number of studies support the notion that FGF23, a bone-derived hormone, not only regulates the most important mineral metabolism (MM) related factors (phosphate, parathyroid hormone, vitamin D, etc.), but might be also involved in cardiovascular (CV) outcome, both in chronic kidney disease (CKD) patients and in the general population. Furthermore, in addition to some direct autocrine and paracrine effects in bone, FGF23 has been suggested to interact with AHSG. In this study we investigated if AHSG is really produced by bone cells, and if its bone production is related and/or controlled by FGF23, using cultured bone cells, according to a new method recently published by our group. Our data show that AHSG is consistently produced in osteocytes and to a far lesser extent in osteoblasts. Both FGF23 addition to the culture medium and its over-expression in osteocytes were associated with a consistent increase of both AHSG mRNA and protein, while FGF23 silencing was followed by opposite effects. Though most of these results were largely affected by the blockage of FGF23 receptors, the role of these receptors in the different experimental sets is still not completely clarified. In addition, we found that FGF23 and AHSG proteins co-localized both in cytoplasm and nucleus, which suggests a possible reciprocal interactivity. Our data not only confirm that AHSG is produced in bone, mainly in osteocytes, but show for the first time that its production is modulated by FGF23. Since both proteins play important roles in the bone and cardiovascular pathology, these results add new pieces to the puzzling relationship between bone and vascular pathology, in particular in CKD patients, prompting future investigations in this field.


The Real-Life Effectiveness and Care Patterns of Type 2 Diabetes Management in Greece.
To investigate the prevalence of hypoglycaemia during sulfonylurea (SU) treatment of type 2 diabetes mellitus (T2DM) in Greece and its influence on glycaemic control, treatment adherence and quality of life (QoL). This was a retrospective cross-sectional study. We included 383 T2DM patients ≥30 years old on treatment with SU in monotherapy or in combination with metformin for at least 6 months. Patients were requested to fill in retrospective questionnaires on hypoglycaemia experience, adherence, weight gain and lifestyle/behavioural factors along with QoL (EQ-5D-3L), treatment satisfaction (TSQM), and fear of hypoglycaemia (HFS-II Worry scale). HbA1c<7% was found in 161 (42.0%) patients. In total, 165 (43.1%) patients reported hypoglycaemic symptoms during the previous 6 months: 41.6% (67/161) of those with HbA1c <7% and 44.1% (98/222) of those with HbA1c ≥7%. Glycaemic control was achieved by 43.1% (94/218) of patients without hypoglycaemia and 50.0% (41/82), 36.8% (25/68) and 6.7% (1/15) of patients with mild, moderate or severe hypoglycaemia, respectively (p=0.013). In multivariate analysis, both occurrence (none vs. mild/moderate/severe) and severity (none vs. mild vs. moderate vs. severe) of hypoglycaemia were significantly associated with impaired global treatment satisfaction (p=0.002 and p<0.0001 respectively) and HFS-II Worry scale scores (both p<0.0001), while lower QoL (EQ-5D (UK) Index) was related to hypoglycaemia severity (p=0.024) only. Finally, treatment adherence was associated with increased (none/mild vs. moderate/severe) hypoglycaemia severity in univariate analysis (p=0.019). A high prevalence of patient treated with SU reported hypoglycaemia in Greek healthcare settings with negative effects on treatment satisfaction, patient worry and adherence. Severity of hypoglycaemic symptoms was associated with reduced glycaemic control.


[Physiological differences and their genetic backgrounds of Erigeron breviscapus populations under enhanced UV-B radiation].
A simulation experiment was conducted under greenhouse condition to study the effects of enhanced UV-B radiation (280-320 nm) on the flavonoid and MDA contents and antioxidant enzyme activities of six Erigeron breviscapus populations, and the intraspecific differences in physiology to the UV-B radiation. The related genetic backgrounds of E. breviscapus populations were also analyzed by ISSR analysis. Under enhanced UV-B radiation, the flavonoid content of E. breviscapus populations D01, D53, D63 and D65 increased significantly at seedling, flowering, and maturing stages, and the MDA content decreased significantly at seedling and flowering stages; while the flavonoid content of D47 and D48 increased significantly at flowering stage but decreased at maturing stage, and the MDA content increased significantly at the three growth stages. The POD and APX activities of D01 at the three growth stages, CAT activities at seedling and flowering stages, and SOD activity at flowering stage significantly increased; the SOD, CAT and APX activities of D47 at the three growth stages and the POD activity at maturing stage decreased significantly; the POD and APX activities of D48 at the three growth stages and the SOD activity at seedling and maturing stages decreased significantly; the SOD and APX activities of D53 at seedling and flowering stages and the CAT activity at flowering stage increased significantly; the SOD, POD and APX activities of D63 at the three growth stages increased significantly; and the CAT and APX activities of D65 at maturing stage had no significant changes, but the test enzyme activities at the three growth stages increased significantly. D01, D53, D63, and D65 were tolerant to UV-B radiation, while D47 and D48 were sensitive to UV-B radiation. The responses of E. breviscapus populations to enhanced UVB radiation varied with growth stage, being in the order of flowering stage > seedling stage > maturing stage. There was an obvious genetic polymorphism among the six E. breviscapus populations, which were divided into two groups at the level of 0.11 of genetic distance, one group including D01, D53, D63 and D65, and the other including D47 and D48. This classification was consistent with the results of physiological response index to UV-B radiation.


Description of a new species of Neoromicia (Chiroptera: Vespertilionidae) from southern Africa: A name for "N. cf. melckorum".
The taxonomy of sub-Saharan small insectivore bats of the family Vespertilionidae is unresolved and currently five named species of the genus Neoromicia are recognized from southern Africa, with N. melckorum considered a synonym of N. capensis. Since several years, the name "N. cf. melckorum" has been used in the literature to designate an apparently undescribed and moderately large bodied vespertilionid bat known from different localities in southern and southeastern Africa. Using new data from molecular genetics, bacular morphology, and cranio-dental characters, we conclude that N. melckorum sensu stricto is indeed nested within N. capensis and obtain the needed evidence to formally describe "N. cf. melckorum", named herein as N. stanleyi sp. nov. On the basis of molecular and bacular evidence, N. stanleyi is found in Botswana, Zimbabwe, and Zambia, and using a combination of other characters is presumed to occur in northern South Africa and Malawi. Bayesian and maximum likelihood analyses based on 12S rRNA sequences indicate that it belongs to a clade containing four species of Neoromicia (N. capensis, N. malagasyensis, N. matroka, and N. robertsi) and Laephotis. Neoromicia stanleyi shows at least 3.2% nucleotide divergence from its closest relatives. It is larger in cranial characters than other members of the capensis group occurring in the southern portion of Africa, and a number of bacular characters distinguish N. stanleyi from N. capensis.


A sensitivity analysis of repeat migration attrition in the study of migrant adjustment: the case of Bangkok.
Studies of migrant adjustment often conclude that results apply only to remaining migrants. This paper examines the potential bias in using the difference between remaining migrants and natives as a measure of migrant adjustment. The results document that differences between remaining migrants and natives contain bias caused by attrition due to repeat migration. Such bias is small, however, and is unlikely to change migrant-native comparisons. Unless one is concerned with details of differences between migrants and natives, it is unnecessary to be concerned about migration attrition bias in drawing conclusions from the observed differences.


Classical counterparts of quantum attractors in generic dissipative systems.
In the context of dissipative systems, we show that for any quantum chaotic attractor a corresponding classical chaotic attractor can always be found. We provide a general way to locate them, rooted in the structure of the parameter space (which is typically bidimensional, accounting for the forcing strength and dissipation parameters). In cases where an approximate pointlike quantum distribution is found, it can be associated with exceptionally large regular structures. Moreover, supposedly anomalous quantum chaotic behavior can be very well reproduced by the classical dynamics plus Gaussian noise of the size of an effective Planck constant ℏ_{eff}. We give support to our conjectures by means of two paradigmatic examples of quantum chaos and transport theory. In particular, a dissipative driven system becomes fundamental in order to extend their validity to generic cases.


Telomerase activity in pancreatic endocrine tumors.
Pancreatic endocrine tumors (PETs) have variable prognoses, and predictors of survival are lacking. PETs can be difficult to distinguish histologically from aggressive pancreatic neoplasms such as acinar cell carcinoma. Telomerase is a ribonuclear protein that maintains the length of the telomere and induces cell immortality. Telomerase is present in 95% of pancreatic adenocarcinoma and is associated with aggressive tumor behavior. Our aim is to determine telomerase activity in PETs and investigate its potential role as a prognostic indicator. Telomerase detection using the telomeric repeat amplification protocol was performed on frozen surgical archived pancreatic endocrine tissue from 30 patients with PETs identified by light microscopy (hematoxylin-eosin stain). All results were confirmed with internal controls. A patient's survival was measured from the time of surgery. Acinar cell differentiation (presence of zymogen granules) was determined by electron microscopy. Follow-up data were acquired via telephone interview, medical record review, and death certificates. Three of 30 PETs diagnosed by light microscopy were telomerase positive: three were considered nonfunctional, and two of these three patients had extrapancreatic disease. All three telomerase-positive cases were reclassified as either acinar cell carcinoma (two cases) or mixed acinar-endocrine cell carcinoma (one case). All three patients (mean age = 63 yr) died from tumor progression within 2 yr of surgery (mean = 1.6 yr +/- 0.5 SD). The remaining PETs were telomerase negative: 13 insulinomas, four nonfunctional, two sporadic glucagonomas, one gastrinoma, one vipoma, one carcinoidlike PET, and five PETs from three patients with multiple endocrine neoplasm syndrome type I and two patients with von Hippel-Lindau syndrome. Excluding insulinomas, 12 of 14 patients with telomerase-negative PETs had extrapancreatic disease. Nevertheless, Kaplan-Meier survival estimates for these 12 patients were significantly longer than for patients with telomerase-positive acinar cell carcinoma (92% vs 0% at 2 yr, p = 0.001, log rank test). The survival of all telomerase-negative PETs (n = 27) was significantly longer than that of the patients with telomerase-positive acinar cell carcinoma (93% vs 0% at 2 yr, p = 0.0001). Telomerase activity helps to identify acinar cell carcinomas that histologically resemble PETs, which accounts for the poor prognosis demonstrated in these patients. The absence of telomerase activity in most PETs may be responsible for their indolent clinical course. Telomerase may identify potentially progressive tumors, such as acinar cell carcinoma, and may be useful in selecting patients for more aggressive treatment.


Group schema therapy for patients with severe anxiety disorders.
This article presents an adaptation of group schema therapy (GST) developed for patients with severe anxiety. An anonymized case presentation exemplifies the adaptation. GST was originally developed to treat clients with maladaptive personality traits but has been applied to a variety of disorders in recent years. For patients with severe anxiety, who have not responded to other treatments, GST may be a promising approach. The paper presents the GST model, the evidence supporting it, and its adaptation for the treatment of anxiety. The case presentation illustrates the therapeutic process and GST interventions. Finally, the clinical outcome and the implications for implementing GST are discussed.


Synthesis Characterization and Photocatalytic Studies of Cobalt Ferrite-Silica-Titania Nanocomposites.
In this work, CoFe₂O₄@SiO₂@TiO₂ core-shell magnetic nanostructures have been prepared by coating of cobalt ferrite nanoparticles with the double SiO₂/TiO₂ layer using metallorganic precursors. The Transmission Electron Microscopy (TEM), Energy Dispersive X-Ray Analysis (EDX), Vibrational Sample Magnetometer (VSM) measurements and Raman spectroscopy results confirm the presence both of the silica and very thin TiO₂ layers. The core-shell nanoparticles have been sintered at 600 °C and used as a catalyst in photo-oxidation reactions of methylene blue under UV light. Despite the additional non-magnetic coatings result in a lower value of the magnetic moment, the particles can still easily be retrieved from reaction mixtures by magnetic separation. This retention of magnetism was of particular importance allowing magnetic recovery and re-use of the catalyst.


[Prospective study of the clinical and urodynamic results of intradetrusor botulinum toxin injections for the treatment of neurogenic overactive bladder].
To evaluate the clinical and urodynamic efficacy and duration of efficacy of intradetrusor botulinum toxin A (BTA) injections in patients with neurogenic overactive bladder refractory to anticholinergic therapy. Between 2004 and 2005, 33 patients (19 men and 14 women), with neurogenic overactive bladder were treated by intradetrusor injections of 300 U of BTA (Botox) in 30 points. All patients were refractory to, intolerant of or presented contraindications to anticholinergic therapy. All patients voided by intermittent self-catheterization. Six weeks after BTA injections, the success rate was 75.8%, with improvement in 12.1% of cases and failure in 9.1% of cases. The mean number of self-catheterizations per 24 h was significantly decreased (6.37 versus 5.2, P=0.02), the maximum voiding volume was increased (321.68 ml versus 536.25 ml, P=0.002), the mean number of episodes of incontinence per 24 h was decreased (7.39 versus 0.03, P<0.0001), the proportion of patients with incontinence was decreased (66.66% versus 6.04%, P<0.0001), the mean maximum cystomanometric capacity was increased (286.75 ml versus 554.16 ml, P=0.002) and the mean maximum intravesical pressure was decreased (54.8 cm H(2)O versus 5.3 cm H(2)O, P<0.0001). After BTA injections, 87.8 % of patients no longer experienced uninhibited contractions. The median duration of clinical efficacy was 7.03 months. At 12 months, injections were still effective clinically in 21.2 % of patients. Intradetrusor BTA injections are an effective and well tolerated treatment for neurogenic overactive bladder. Their clinical efficacy persisted for more than 12 months in more than 20% of cases.


Squamous neoplasia of vagina related to DES syndrome.
The emphasis to date in the DES syndrome has been confined to the presence of adenosis in the vagina and the occassional adenocarcinoma which may arise from this ectopic glandular epithelium (less than 250 to date). As these patients mature, however, the glandular epithelium in the vagina is replaced by squamous metaplastic epithelium. Forty-six patients with benign vaginal adenois were observed colposcopically for up to 2 years. Atypical squamous epithelium (white epithelium, punctation, or mosiac pattern) was observed in 40 of the 46 patients. Colposcopically directed biopsy revealed six cases ofdysplasia (mild to severe) and one unequivocal case of in situ squamous carcinoma. These squamous lesions in the vagina were identical, both colposcopically and histologically,to similiar lesions on the cervix. These data suggest that squamous neoplasia may be more frequent in the future of the DES-exposed patient than adenocarcinoma.


The mutagenic effect of pesticides on Escherichia coli WP2 try.
Thirty pesticides commercially available in Hungary and three well-known chemical mutagens were applied to agar plate cultures of her+ and her- derivatives of Escherichia coli WP2 try- strain. After incubation, the plates were examined for a relative increase in reverse mutation number. N-Trichloro methylthio-1,2,3,6-tetrahydrophthalimide, dimethyl-3,2-dichlorovinyl-phosphate and N-trichloro methylthiophyhalamide proved to have definite mutagenic activity.


Sensitive determination of triacetone triperoxide explosives using electrogenerated chemiluminescence.
Sensitive and selective detection and quantification of high explosive triacetone triperoxide (TATP) with electrogenerated chemiluminescence (ECL) at a glassy carbon electrode in water-acetonitrile solvent mixture were reported. In the presence of ruthenium(II) tris(bipyridine), TATP or hydrogen peroxide derived from TATP via UV irradiation or acid treatment produced ECL emissions upon cathodic potential scanning. Interference from hydrogen peroxide on TATP detection was eliminated by pretreatment of the analyte with catalase enzyme. Selective detection of TATP from hexamethylene triperoxide diamine (HMTD, another common peroxide-based explosive) was realized by comparing ECL responses obtained from the anodic and the cathodic potential scanning; TATP produced ECL upon cathodic potential scanning only, whereas HMTD produced ECL upon both cathodic and anodic potential scanning. The hydroxyl radical formed after the electrochemical reduction of TATP was believed to be the key intermediate for ECL production, and its stability was strongly dependent on the solution composition, which was verified with electron paramagnetic resonance spectroscopy. A detection limit of 2.5 μM TATP was obtained from direct electrochemical reduction of the explosive or hydrogen peroxide derived from TATP in 70/30% (v/v) water-acetonitrile solutions, which was ~400 times lower than that reported previously based on liquid chromatography separation and Fourier transform infrared detection.


Serum lipoproteins and apolipoproteins in young male survivors of myocardial infarction.
Concentrations of serum lipoprotein lipids and apolipoproteins A-I, A-II and B were determined 3-6 months after myocardial infarction in 116 males below the age of 45 and in 116 age-matched controls. Among single variables the sum of cholesterol concentration in VLDL and LDL divided by the HDL cholesterol level was the best discriminator between patients and controls. The concentrations of serum triglycerides, apolipoprotein B, VLDL triglycerides and cholesterol, serum cholesterol, HDL cholesterol and LDL triglycerides, in that order, were better discriminators than was LDL cholesterol level. Among variables reflecting HDL concentration and composition HDL cholesterol was the best discriminator followed by HDL2 cholesterol, apolipoprotein A-I and the HDL cholesterol/apolipoprotein A-I ratio. Multivariate analysis indicated independent significance of elevated VLDL lipid and LDL cholesterol concentrations, and a decreased HDL cholesterol concentration, in relation to MI. The present data suggest that a disturbed triglyceride metabolism, in addition to elevated LDL and decreased HDL cholesterol levels, has an independent and pathogenetic significance for MI at a young age.


Characterization of two novel cryptococcal mannoproteins recognized by immune sera.
Host defenses against the encapsulated yeast Cryptococcus neoformans involve both humoral and cell-mediated immunity. Mannoproteins (MPs) are a heterogeneous class of immunodominant glycoproteins which have been only incompletely characterized. In this study, we report on the molecular features of two novel MPs that are recognized by serum antibodies during cryptococcosis. After fractionation of extracellular cryptococcal products, MPs reacted more strongly than other components with sera from C. neoformans-infected AIDS patients. Further fractionation and Western blot analysis of MPs evidenced the presence of highly reactive bands with molecular masses of 250, 125, 115, and 84 kDa. The 115- and 84-kDa bands contained significant amounts of N-linked oligosaccharides, as shown by decreased molecular mass after peptide-N-glycosidase F treatment. N-terminal amino acid sequences of the two bands were used to search C. neoformans nucleotide databases. Homologous genomic sequences were used to synthesize DNA probes and isolate cDNA clones containing the full-length genes, which were designated MP84 and MP115. Both genes showed the presence of a serine/threonine-rich region, a potential site for heavy glycosylation. MP84 and MP115 showed homology with, respectively, polysaccharide deacetylases and carboxylesterases from other organisms. Recombinant, deglycosylated proteins expressed in Escherichia coli still reacted with sera from patients, albeit more weakly than natural MPs, indicating that at least some of the reactive epitopes were retained in the recombinant forms. In conclusion, we identified two novel MPs that are important targets of antibody responses during cryptococcosis. These data may be useful to devise alternative immunity-based strategies to control the disease.


Antimicrobial susceptibility pattern of clinical isolates of Burkholderia pseudomallei in Bangladesh.
Melioidosis an infectious disease, caused by a Gram negative bacterium called Burkholderia pseudomallei, is endemic in Bangladesh. This organism is sensitive to limited number of antimicrobial agents and need prolonged treatment. There is no comprehensive data on the antimicrobial susceptibility profile of B. pseudomallei isolated in Bangladesh over last several years. The present study aimed to determine the antimicrobial susceptibility pattern of B. pseudomallei isolated in a tertiary care hospital of Dhaka city from 2009 to 2015. All B. pseudomallei isolated from melioidosis patients over a period of 7 years (2009-2015) in the Department of Microbiology of a 725-bed tertiary care referral hospital in Dhaka city, Bangladesh were included in the study. B. pseudomallei was identified by Gram stain, culture, specific biochemical tests, serology and PCR using specific primers constructed from 16s rRNA region of B. pseudomallei. Antimicrobial susceptibility to specific agents was determined by disk diffusion and minimum inhibitory concentration methods. A total of 20 isolates of B. pseudomallei which were isolated from patients coming from different geographic locations of Bangladesh were included in the study. All the isolates were uniformly sensitive (100%) to ceftazidime, imipenem, piperacillin-tazobactam, amoxicillin-clavulanic acid and tetracycline by both disk diffusion and MIC methods. Two strains were resistant to trimethoprim-sulfamethoxazole by disk diffusion method but were sensitive by MIC method. The MIC50 and MIC90 values of the above antimicrobial agents were almost similar. All the isolates were resistant to amikacin by both MIC and disk diffusion methods. The results of the study suggest that B. pseudomallei prevalent in Bangladesh were still susceptible to all recommended antimicrobial agents used for the treatment of melioidosis. However, regular monitoring is needed to detect any emergence of resistance and shifting of MIC50 and MIC90 values.


Respiration audiometry.
Since 1971 an impedance-plethysmographic method was been used to record changes in the breathing pattern of 218 high-risk infants during acoustic stimulation. The results of this technique, known as respiration audiometry, show that acoustically induced changes in the breathing pattern can be used as a hearing test for very young children. The results fall into four categories: excellent readability (45%), good readability (32%), readable (15%), not readable (8%). Respiration audiometry makes it possible in most cases to ascertain a child's hearing level. The neonatal period and early infancy are the ages best suited for investigations with respiration audiometry. The reliability of this method was found to be good.


Impact of direct drug delivery via gastric access devices.
Gastric access devices such as nasogastric tubes and gastrostomy tubes are increasingly being used in clinical practice to provide both short- and long-term nutrition support therapy. Increasingly these devices are being utilized to help deliver oral medications, where swallowing is impaired. This concomitant administration of medications and enteral formulas could derive potential benefits in regard to time and cost; however, uncertainty exists regarding potential drug and nutrient interactions and the influence this may have on both safety and efficacy. This article provides an overview of the differing gastric access devices used in clinical practice and evaluates the evidence base for using oral medications via these routes. Alternative methods of drug administration are discussed, alongside common drug nutrient interactions and potential complications. Delivering medications via gastric access devices can be performed safely; however, careful consideration needs to be made regarding tube and patient influences, alongside drug-nutrient interactions. Improving practice in this area in the future necessitates enhancement of an evidence base to substantiate the safety of drug delivery via gastric access devices and improvement in education among healthcare professionals about the potential problems.


[Measurement of the pressure in dental casting].
The tests showed that the pressure exerted on the tooth stump and the surrounding tissue is determined by the consistency of the impression material, the amount escaping from the impression tray, the adaptation pressure and adaptation speed. The original consistency of the impression material without exposure to a catalyst is an especially important factor. If the material has a flow limit (e.g. Optosil with or without catalyst) it is exerting pressure in any case and this pressure continues even after setting. Impression materials without flow limit (e.g. Silaplast without catalyst) produce practically no pressure. Only by the activity of the catalyst a flow limit develops and thus pressure is produced.


[The results of combined acupuncture and laser therapy of outpatients with duodenal ulcer].
Exacerbations of duodenal ulcer (DU) were treated outpatient in 130 DU sufferers who failed previous chemotherapy. The treatment consisted of acupuncture, laser therapy, diet and drugs. All the patients achieved remission, relief of vegetative disorders. DU healing took less time (by 7-10 days) than in control group. Application of the acupuncture and laser radiation allowed assignment of minimal drug doses. 3-year long-term results showed the absence of recurrences for 1-3 years. 87 DU patients gave up smoking after additional acupuncture according to the special regimen. The conclusion is made that acupuncture combined with laser therapy is effective against DU, safe and usable in outpatient setting.


Garlic, Onion, and Cinnamon Essential Oil Anti-biofilms' Effect against Listeria monocytogenes.
Biofilms represent a serious problem for food industries due to their persistence in processing surfaces, from which they can cause food spoilage or, even worse, lead to foodborne diseases. Microorganisms immersed in biofilms are more resistant to biocides. The search for natural effective alternatives for the prevention and the control of biofilms has increased lately. The aim of this research was to test the antibacterial and the anti-biofilm activities of cinnamon, onion, and garlic essential oils against Listeria monocytogenes. The methodology highlighted first the effect of these essential oils on L. monocytogenes using disc diffusion and minimum inhibitory concentration (MIC) methods and then on initial cell attachment and six hours preformed biofilms. The inhibition of biofilms was assessed by crystal violet assay. Sulfides were the most abundant compounds present in onion and garlic essential oils, while cinnamaldehyde was predominant in cinnamon essential oil. MIC values were of 0.025 mg mL-1 for onion essential oil and 0.100 mg mL-1 for cinnamon and garlic. Onion essential oil inhibited initial cell attachment by 77% at 0.5 of the MIC dose, while at MIC, cinnamon and garlic essential oils inhibited the initial microbial adhesion completely. All three essential oils completely inhibited initial cell attachment when applied at 2 MIC. On the contrary, preformed biofilms were more resistant, and the inhibition rate ranged from 33% to 78%. In summary, this investigation revealed that the essential oils of garlic, onion, and cinnamon show an effective antibiofilm activity against L. monocytogenes and are promising natural antimicrobial alternatives for food processing facilities.


Association Between Serum Albumin Levels and Obesity and Risk of Developing Chronic Kidney Disease Using Data from the Korean Multi-Rural Communities Cohort (MRCohort) Population Database.
BACKGROUND The Multi-Rural Communities Cohort (MRCohort) was established as part of the Korean Genome and Epidemiology Study, which identified risk factors for cardiovascular diseases in the population between January 2005 and December 2011. This was a prospective study, aimed to investigate the association between serum albumin levels and the risk of developing chronic kidney disease (CKD) in obese patients using data from the MRCohort population database. MATERIAL AND METHODS For analysis, we selected a total of 9695 people from the MRCohort from 2004 to 2013: 3105 people from Goryeong, 3183 from Yangpyeong, and 3407 from Namwon, without previous history of myocardial infarction, cerebrovascular disease, cancer, and CKD, or missing values on confounders. As a result, 2300 metabolically healthy participants were studied as a whole, and then by body mass index of >23 kg/m². The groups were divided into 4 subgroups by albumin quartiles, and the effects of albumin levels compared to the lowest quartile were studied by Cox proportional hazard analysis, after adjusting for potential confounders. RESULTS The results showed that the highest quartile of albumin, compared to the lowest quartile, was a protective factor of CKD development only in the overweight and obese group with adjusted hazard ratio (HR)=0.38 (95% confidence interval (CI) 0.16-0.95, p<0.05). In the normal-weight group, results were insignificant, with adjusted HR=0.54 (95% CI 0.24-1.25, not significant). CONCLUSIONS Data from the Korean MRCohort population database showed that obesity and increased serum albumin levels were protective factors for the development of CKD.


[Correlation of sperm nuclear maturity with the outcomes of in vitro fertilization].
To investigate the correlation between sperm nuclear maturity and the outcomes of in vitro fertilization (IVF). We retrospectively analyzed the clinical data on 1 393 cycles of IVF in our center from September 2018 to May 2020. According to the high DNA stainability (HDS) of sperm, we divided the patients into a mature (HDS ≤ 10%) and an immature sperm nucleus group (HDS > 10%), compared the results of IVF, embryonic development and clinical outcomes between the two groups, and analyzed their correlation with HDS. The rates of fertilization and normal fertilization were significantly higher and that of fertilization failure remarkably lower in the mature than in the immature sperm nuclear group (P < 0.05). There were no statistically significant differences between the two groups in the rates of cleavage, high-quality embryo, blastocyst formation, clinical pregnancy, miscarriage and live birth (P > 0.05). Univariate logistic regression analysis showed that HDS was correlated negatively with the rates of fertilization, normal fertilization and blastocyst formation (P < 0.05), positively with fertilization failure (P < 0.05), but not significantly with the rates of cleavage, high-quality embryo, clinical pregnancy, miscarriage and live birth (P > 0.05). After adjusting for confounding factors by multivariate logistic regression analysis, HDS was found correlated negatively with the fertilization rate (OR = 0.945, 95% CI: 0.918 to 0.972, P < 0.05), positively with fertilization failure (OR = 1.043, 95% CI: 1.010 to -1.078, P < 0.05), but not significantly with the rates of normal fertilization, cleavage, high-quality embryo, blastocyst formation, clinical pregnancy, miscarriage and live birth (P > 0.05). Sperm nuclear maturity significantly affects the rates of fertilization and fertilization failure in IVF patients. Taking sperm nuclear maturity indicators in consideration in the process of IVF may contribute to comprehensive evaluation of semen and male fertility and guide clinical decision-making.


Conclusions from clinical trials of the Southwest Oncology Group.
Mature data from four clinical trials conducted by the Southwest Oncology Group from 1971 to 1978 for patients with all stages of Hodgkin's disease (HD) are reviewed in this paper. In the RAC #1 trial of stage I and II HD we demonstrated that involved-field radiotherapy plus six courses of MOPP chemotherapy improved relapse-free survival compared to standard radiotherapy alone (P = 0.12), especially in patients with B symptoms (P less than 0.03) or mediastinal disease (P = 0.08). However, at present, there is no significant difference in overall survival. In the CAR #1 study for patients with pathologic stage IIB, IIIA, and IIIB HD, we demonstrated that three or four courses of MOPP before radiotherapy produced a 90% complete remission (CR) rate, with 70% of the patients remaining free of disease at 5 years. In the CAR #2 study for patients with pathologic stage IIIA or IIIB disease, we demonstrated that chemotherapy alone (MOPP-bleomycin) was as effective as combined modality treatment (MOPP-bleomycin plus radiotherapy) in terms of CR rate (85% versus 89%, respectively), relapse-free survival, and survival. For advanced stages of HD we added doxorubicin to our MOPP-bleomycin schedule and demonstrated that MOP-BAP produced a 77% CR rate compared to 67% for MOPP-bleomycin (P = 0.10). Moreover, MOP-BAP produced consistently superior CR rates and survival in patients with more prognostically favorable presentations of HD. Our new ongoing study (MOPP #6) incorporates many of the concepts derived from these earlier clinical investigations.


Identification of enterotoxigenic Escherichia coli isolated from swine with diarrhea in Thailand by colony hybridization, using three enterotoxin gene probes.
The DNA colony hybridization assay was used to identify enterotoxigenic Escherichia coli among E. coli isolated from 803 swine with diarrhea at 10 farms in Thailand. Between 5 September and 8 December 1981, enterotoxigenic E. coli were identified in 40% of 58 litters of piglets under 10 days old and 17% of 29 litters between 10 and 21 days old with diarrhea at farms at four different locations in Thailand. All E. coli that hybridized with one or more of the three enterotoxin gene probes produced heat-labile or heat-stable toxin or both, as determined by testing culture supernatants in the Y1 adrenal and suckling mouse assays. The DNA colony hybridization technique is a specific method of identifying enterotoxigenic E. coli from swine and can be used to further characterize these enteric pathogens.


Factors influencing health behaviors in the National Health and Nutritional Examination Survey, III (NHANES III).
This study investigated the influence of age, gender, race, place of residence, social networks, and socioeconomic status (SES) on health behaviors in the NHANES III, a large public domain database of approximately 16,000 subjects. Multiple regression analysis indicated that age, gender, social networks, and SES were statistically significant predictors of both positive and negative health behaviors, while race and place of residence were not. These results suggest an influence of age, gender, SES, and social support factors on health behaviors and reinforce the need for social work to take into account these factors at both the individual and public policy levels.


Image Distortion in Biplanar Slot Scanning: Part 2 Technology-specific Factors.
Biplanar digital slot scanning technology has become the standard of care in the treatment of scoliosis. Yet, the amount of distortion and reproducibility of this type of imaging modality has yet to be fully investigated. In our paper "Image distortion in biplanar slot scanning: part 1 patient-specific factors" we found that there was potentially clinically impactful interimage distortion. The purpose of this study was to evaluate the degree to which this image distortion was secondary to the image acquisition process. Four 25 mm radio-opaque markers were placed at C3, T1, T12, and L5 on a full-length skeleton model. The skeleton was imaged in 10 different positions within the scanner. Five posteroanterior and 5 lateral images were obtained in each position. Two orthopaedic attending physicians and 3 orthopaedic resident physicians measured the markers for a total of 3200 measurements. Intraclass correlation coefficients (ICCs) and 95% confidence intervals were used to examine image distortion. Average marker size was 24.77, with a standard error of measurement of 0.00493. Image distortion and standard error of measurement accounted for ∼0.5% to 1.5% of total the measurement. Overall, there was good reliability and consistency when looking at markers in different views (ICC 0.790), planes, and locations within the image. Horizontal measurements were found to be more consistent and have better reliability (ICC 0.881) than vertical measurements (ICC 0.386). Position within the scanner had minimal impact on the accuracy of the measurements. This study demonstrates that there is minimal error due to image acquisition and measurement when using a biplanar slot scanner. Biplanar slot scanning technology tended to underestimate the size of the marker; however, the least accurate measurements only erred by 1.5% from the true length. This indicates that unlike traditional radiographs the sources of error in biplanar slot scanning images are not due to parallax and are likely due to patient-specific factors and rather than the technology itself.


Toward Rational Design of Novel Anti-Cancer Drugs Based on Targeting, Solubility, and Bioavailability Exemplified by 1,3,4-Thiadiazole Derivatives Synthesized Under Solvent-Free Conditions.
The 1,3,4-thiadiazole derivatives (9a-i) were synthesized under solvent free conditions and their chemical composition was confirmed using different spectral tools (IR, Mass, and NMR spectrometry). All the synthesized compounds were screened for their anti-cancer potentiality over human breast carcinoma (MCF-7) and human lung carcinoma (A-549). Most of the tested compounds showed remarkable anti-breast cancer activity. However, compound 4 showed the most anti-lung cancer activity. Then, compounds with cytotoxic activity ≥ 80% over breast and lung cells were subjected to investigate their specificity on human normal skin cell line (BJ-1). Compounds 9b and 9g were chosen owing to their high breast anti-cancer efficacy and their safety, in order to study the possible anti-cancer mode of action. Otherwise, drug delivery provides a means to overcome the low solubility, un-targeted release, and limited bioavailability of the prepared 1,3,4-thiadiazole drug-like substances. Compounds 9b and 9g were chosen to be encapsulated in Na-alginate microspheres. The release profile and mechanism of both compounds were investigated, and the results revealed that the release profiles of both microspheres showed a sustained release, and the release mechanism was controlled by Fickian diffusion. Accordingly, these compounds are promising for their use in chemotherapy for cancer treatment, and their hydrophilicity was improved by polymer encapsulation to become more effective in their pharmaceutical application.


Ecological risk assessment of heavy metals in sediment and human health risk assessment of heavy metals in fishes in the middle and lower reaches of the Yangtze River basin.
The concentrations of heavy metals (Cr, Cd, Hg, Cu, Zn, Pb and As) in the water, sediment, and fish were investigated in the middle and lower reaches of the Yangtze River, China. Potential ecological risk analysis of sediment heavy metal concentrations indicated that six sites in the middle reach, half of the sites in the lower reach, and two sites in lakes, posed moderate or considerable ecological risk. Health risk analysis of individual heavy metals in fish tissue indicated safe levels for the general population and for fisherman but, in combination, there was a possible risk in terms of total target hazard quotients. Correlation analysis and PCA found that heavy metals (Hg, Cd, Pb, Cr, Cu, and Zn) may be mainly derived from metal processing, electroplating industries, industrial wastewater, and domestic sewage. Hg may also originate from coal combustion. Significant positive correlations between TN and As were observed.


Mitochondrial aconitase modification, functional inhibition, and evidence for a supramolecular complex of the TCA cycle by the renal toxicant S-(1,1,2,2-tetrafluoroethyl)-L-cysteine.
Metabolism of the common industrial gas tetrafluoroethylene in mammals results in the formation of S-(1,1,2,2)-tetrafluoroethyl-L-cysteine (TFEC), which can be bioactivated by a mitochondrial C-S lyase commonly referred to as beta-lyase. The resultant "reactive intermediate", difluorothioacetyl fluoride (DFTAF), is a potent thioalkylating and protein-modifying species. Previously, we have identified mitochondrial HSP70, HSP60, aspartate aminotransferase, and the E2 and E3 subunits of the alpha-ketoglutarate dehydrogenase (alphaKGDH) complex as specific proteins structurally modified during this process. Moreover, functional alterations to the alphaKGDH complex were also detected and implicated in the progression of injury. We report here the identification, by tandem mass spectrometry, and functional characterization of the final remaining major protein species modified by DFTAF, previously designated as P99(unk), as mitochondrial aconitase. Aconitase activity was maximally inhibited by 56.5% in renal homogenates after a 6 h exposure to TFEC. In comparison to alphaKGDH, aconitase inhibition (up to 79%) in a cell culture model for TFEC-mediated cytotoxicity was greater and preceded alphaKGDH inhibition, indicating that aconitase modification may constitute an early event in TFEC-mediated mitochondrial damage and cell death. These findings largely define the initial lesion of TFEC-mediated cell death and also have implications for the modeling of mitochondrial enzymatic architecture and the localization and identity of renal mitochondrial cysteine S-conjugate beta-lyase.


Conformational alteration of mRNA structure and the posttranscriptional regulation of erythromycin-induced drug resistance.
The DNA sequence of the ermC gene of plasmid pE194 is presented. This determinant is responsible for erythromycin-induced resistance to the macrolide-lincosamide-streptogramin B group of antibiotics and specifies a 29,000 dalton inducible protein. The locations of the ermC promoter, as well as that of a probable transcriptional terminator, are established both from the sequence and by transcription mapping. The sequence contains an open reading frame sufficient to encode the previously identified 29,000 dalton ermC protein. Between the promoter and the putative ATG start codon is a 141 base pair leader sequence, within which several regulatory (constitutive) mutations have been mapped and sequenced. The leader has a second open reading frame, sufficient to encode a 19 amino acid peptide. It is suggested that induction by erythromycin involves a shift between alternative ribosome-bound mRNA conformations, so that the ribosome binding sequence and the start codon for synthesis of the 29K protein are unmasked in the presence of inducer. Possible active and inactive folded configuration of the leader sequence are presented, as well as the effects on these configurations of regulatory mutations.


[New functional proteins identified by proteomic analysis in the epidermal growth factor receptor-mediated signaling pathway and application for practical use].
To clarify the whole picture of epidermal growth factor (EGF) signaling pathway, we identified proteins from the EGF-stimulated A431 cells by anti-phospho-tyrosine antibody column chromatography. Over 150 proteins were detected including previously unidentified proteins as well as well-studied proteins. Among these proteins, we picked up four proteins that had not been known in EGF signaling pathway and analyzed their functions. We report the functions of these proteins in this article. 1) CFBP interacts with CD2AP family proteins and functions as a key component in downregulation of EGF receptor protein level following EGF stimulation. 2) Ymer is found to be phosphorylated and ubiquitinated upon EGF stimulation, and functions as a regulator for the downregulation and endocytosis of EGF receptor. 3) CLPABP binds to mitochondria-specific phospholipids, cardiolipin, through its PH domain, and its complex includes various proteins related to mRNA metabolism. 4) GAREM is associated with Grb2 and Shp2. Each association affects the ERK activity. Finally, we discuss the possibilities that these proteins can be used as a novel biomarker protein for cancer and other diseases.


Impact of γ'γ' fibrinogen interaction with red blood cells on fibrin clots.
γ' fibrinogen has been associated with thrombosis. Here the interactions between γ'γ' or γAγA fibrinogen and red blood cells (RBCs), and their role on fibrin clot properties were studied. Atomic Force microscopy (AFM)-based force spectroscopy, rheological, electron and confocal microscopy, and computational approaches were conducted for both fibrinogen variants. AFM shows that the recombinant human (rh)γ'γ' fibrinogen increases the binding force and the frequency of the binding to RBCs compared with rhγAγA, promoting cell aggregation. Structural changes in rhγ'γ' fibrin clots, displaying a nonuniform fibrin network were shown by microscopy approaches. The presence of RBCs decreases the fibrinolysis rate and increases viscosity of rhγ'γ' fibrin clots. The full length of the γ' chain structure, revealed by computational analysis, occupies a much wider surface and is more flexible, allowing an increase of the binding between γ' fibers, and eventually with RBCs.


Cloning and characterization of a tyrosinase gene from the white-rot fungus Pycnoporus sanguineus, and overproduction of the recombinant protein in Aspergillus niger.
A new tyrosinase-encoding gene (2,204 bp) and the corresponding cDNA (1,857 nucleotides) from the white-rot fungus Pycnoporus sanguineus BRFM49 were cloned. This gene consisted of seven exons and six introns and encoded a predicted protein of 68 kDa, exceeding the mature tyrosinase by 23 kDa. P. sanguineus tyrosinase cDNA was over-expressed in Aspergillus niger, a particularly suitable fungus for heterologous expression of proteins of biotechnological interest, under the control of the glyceraldehyde-3-phosphate-dehydrogenase promoter as strong and constitutive promoter. The glucoamylase preprosequence of A. niger was used to target the secretion. This construction enabled the production of recombinant tyrosinase in the extracellular medium of A. niger. The identity of the purified recombinant protein was confirmed by N-terminal amino acid sequencing. The maturation process was shown to be effective in A. niger, and the recombinant enzyme was fully active, with a molecular mass of 45 kDa. The best transformant obtained, A. niger D15#26-e, produced extracellular tyrosinase activities of 534 and 1,668 U l(-1) for monophenolase and diphenolase, respectively, which corresponded to a protein yield of ca. 20 mg l(-1).


Contact activation via ICAM-1 induces changes in airway epithelial permeability in vitro.
The role of ICAM-1 in contact activation of the bronchial epithelial cells is elucidated. Direct contact between epithelial cells and leukocytes is required to change transepithelial electrical resistance (TER) of the epithelium. Migration of human neutrophils across the layers of cultured human airway epithelial cells (Calu-3) or primary cow tracheal epithelial cells was induced by an fMLP gradient. Migrating neutrophils decreased TER and increased permeability to albumin. Monoclonal antibodies to ICAM-1 reduced neutrophil migration, thus reducing the changes in TER and changes in the epithelial permeability to albumin. By confocal microscopy, ERK1/2 was found to be locally activated in the epithelial cells at the sites of migration and cross-linking of ICAM-1. Blockade of ERK1/2 by PD98059 decreased the changes in TER which were induced by ICAM-1 cross-linking. Contact activation of the bronchial epithelial cells, involving ICAM-1 via local activation of ERK1/2, is an important mechanism of alteration of the bronchial epithelial permeability.


Facing hierarchy: a qualitative study of residents' experiences in an obstetrical simulation scenario.
Residents in surgical specialties face a steep hierarchy when managing medical crises. Hierarchy can negatively impact patient safety when team members are reluctant to speak up. Yet, simulation has scarcely been previously utilized to qualitatively explore the way residents in surgical specialities navigate this challenge. The study aimed to explore the experiences of residents in one surgical specialty, obstetrics and gynecology (Ob/Gyn), when challenging hierarchy, with the goal of informing future interventions to optimize resident learning and patient safety. Eight 3rd- and 4th-year Ob/Gyn residents participated in a simulation scenario in which their supervising physician made an erroneous medical decision that jeopardized the wellbeing of the labouring mother and her foetus. Residents participated in 30-45 min semi-structured interviews that explored their approach to managing this scenario. Transcribed interviews were analysed using qualitative thematic inquiry by three research team members, finalizing the identified themes once consensus was reached. Study results show that the simulated scenario did create an experience of hierarchy that challenged residents. In response, residents adopted three distinct communication strategies while confronting hierarchy: (1) messaging - a mere reporting of existing clinical information; (2) interpretive - a deliberate construction of clinical facts aimed at swaying supervising physician's clinical decision; and (3) advocative - a readiness to confront the staff physician's clinical decision. Furthermore, residents utilized coping mechanisms to mitigate challenges related to confronting hierarchy, namely deflecting responsibility, diminishing urgency, and drafting allies. Both these communication strategies and coping mechanisms shaped their practice when challenging hierarchy to preserve patient safety. Understanding the complex processes in which residents engage when confronting hierarchy can serve to inform the development and study of curricular innovations. Informed by these processes, we must move beyond solely teaching residents to speak up and consider a broader curriculum that targets not only residents but also faculty physicians and the learning environment within the organization.


Chromosome comparison among five species of Neotropical cichlids of Cichlasoma and Gymnogeophagus (Perciformes).
The genera Cichlasoma and Gymnogeophagus belong to the subfamily Cichlinae, the only one in Neotropical cichlids. Cichlasoma dimerus, C. paranaense, C. portalegrense, Gymnogeophagus rhabdotus, and G. lacustris were collected at different points in the Paranapanema and Paraguay basins and the Lagoon of Patos hydrographic system. In addition to conventional analysis, CMA3 fluorochrome staining, and FISH with 18S rDNA probe were performed. All species had a diploid number equal to 48, with interand intraspecific differences in karyotype formulae. All species presented a single AgNOR site, except G. rhabdotus and the C. paranaense population of the Paranapanema River, which revealed more than one pair of nucleolar chromosomes. AgNORs were coincident to 18S rDNA and CMA3. Heterochromatin was distributed in the pericentromeric chromosomal regions and coincident with NORs. For the first time, this work shows cytogenetic data for C. portalegrense, G. lacustris, and G. rhabdotus. Although some results reinforce the idea of conservative chromosome evolution of 2n in Cichlinae, interspecific and populational variations observed confirm that chromosomal rearrangements affect the microstructural karyotype diversification in this group of fish.