Miracle-dz/UniFAR-FTUs huggingface dataset:
Sau3AI, a Monomeric Type II Restriction Endonuclease That Dimerizes on the DNA and Thereby Induces DNA Loops*

Here, we report that Sau3AI, an unusually large type II restriction enzyme with sequence homology to the mismatch repair protein MutH, is a monomeric enzyme as shown by gel filtration and ultracentrifugation. Structural similarities in the N- and C-terminal halves of the protein suggest that Sau3AI is a pseudo-dimer, i.e. a polypeptide with two similar domains. Since Sau3AI displays a nonlinear dependence of cleavage activity on enzyme concentration and a strong preference for substrates with two recognition sites over those with only one, it is likely that the functionally active form of Sau3AI is a dimer of a pseudo-dimer. Indeed, electron microscopy studies demonstrate that two distant recognition sites are brought together through DNA looping induced by the simultaneous binding of twoSau3AI molecules to the DNA. We suggest that the dimeric form of Sau3AI supplies two DNA-binding sites, one that is associated with the catalytic center and one that serves as an effector site.
Circadian chemotherapy for gynecological and genitourinary cancers

The circadian timing of surgery, anticancer drugs, radiation therapy, and biologic agents can result in improved toxicity profiles, tumor control, and host survival. Optimally timed cancer chemotherapy with doxorubicin or pirarubicin (06:00h) and cisplatin (18:00h) enhanced the control of advanced ovarian cancer while minimizing side effects, and increased the response rate in metastatic endometrial cancer. Therapy of metastatic bladder cancer with doxorubicin–cisplatin was made more tolerable by this same circadian approach resulting in a 57% objective response rate. This optimally timed therapy is also effective in the adjuvant setting, decreasing the expected frequency of metastasis from locally advanced bladder cancer. Circadian fluorodeoxyuridine (FUDR) continuous infusion (70% of the daily dose given between 15:00h and 21:00h) has been shown effective for metastatic renal cell carcinoma resulting in 29% objective response and stable disease of more than 1 yr duration in the majority of patients. Toxicity is reduced markedly when FUDR infusion is modulated to circadian rhythms. In a multicenter trial in patients with metastatic renal cell cancer, patients were randomized to a flat or a circadian-modified FUDR infusion. This study confirmed a significant difference in toxicity and dose intensity, favoring the circadian-modified group. Hormone refractory metastatic prostate cancer has been treated with circadian-timed FUDR chemotherapy; however, without objective response. Biological agents such as interferon-α and IL-2 have shown low but effective disease control in metastatic renal cell cancer, however, with much toxicity. Each of these cytokines shows circadian stage dependent toxicity and efficacy in model systems. In summary, the timing of anthracycline, platinum, and fluoropyrimidine-based drug therapies during the 24h is relevant to the toxic–therapeutic ratio of these agents in the treatment of gynecologic and genitourinary cancers.
Introducing undergraduate medical teaching into general practice: an action research study

Following the publication of Tomorrow's Doctors and as a result of increasing numbers of students recruited to medical school it is necessary to involve more general practitioners (family physicians) in undergraduate medical education. Students have responded positively regarding experiences in general practices with a broad spectrum of clinical conditions to be seen and greater involvement in clinical decision-making. This action research study followed a small group general practice in South Wales through the required preparation for undergraduate medical education and its first year of teaching. Preparatory work for the practice focused mainly on summarizing patient notes, setting up a practice library and arranging accommodation for the students. Members of the Primary Health Care Team (PHCT) found that having students in the practice gave them a sense of achievement and enhanced self-worth. Individuals within the practice felt more confident in their professional role and the team ethic within the practice was strengthened. Doctors’ anxieties regarding the adequacy of their clinical skills proved unfounded. Patients were reported to feel more included in their care and to have enjoyed hearing their condition being discussed with the students. Students valued the one-to-one teaching, seeing common illnesses and a variety of consulting styles. It is hoped that this paper will be of value to those responsible for recruiting GP practices into undergraduate teaching. It demonstrates benefits for the primary health care team in terms of improved morale and sense of professional self-worth. Patients felt more involved in their care. Generalization from these findings is limited by only one practice having been involved. Undergraduate teaching offers advantages, particularly in terms of professional self-esteem and team morale. Practice points•Having medical students in the practice gave members of the primary health care team an enhanced sense of self-worth and made them feel more confident in their professional role. •The team ethic within the practice was strengthened by the presence of the students. •Doctors were anxious about the adequacy of their clinical skills before the students started but this proved unfounded. •Reports suggested that patients felt more included in their care and enjoyed hearing their condition being discussed with the students.
Insulin pump therapy: a meta-analysis.

OBJECTIVE To conduct a meta-analysis of the metabolic and psychosocial impact of continuous subcutaneous insulin infusion (CSII) therapy on adults, adolescents, and children. RESEARCH DESIGN AND METHODS Studies were identified and data regarding study design, year of publication, sample size, patient's age, diabetes duration, and duration of CSII therapy were collected. Means and SDs for glycohemoglobin, blood glucose, insulin dosages, and body weight for CSII and comparison conditions were subjected to meta-analytic procedures. Data regarding pump complications and psychosocial functioning were reviewed descriptively. RESULTS A total of 52 studies, consisting of 1,547 patients, were included in the meta-analysis. Results indicate that CSII therapy is associated with significant improvements in glycemic control (decreased glycohemoglobin and mean blood glucose). A descriptive review of potential complications of CSII use (e.g., hypoglycemia, diabetic ketoacidosis [DKA], pump malfunction, and site infections) suggests a decreased frequency of hypoglycemic episodes but an increased frequency of DKA in studies published before 1993. CONCLUSIONS CSII therapy is associated with improved glycemic control compared with traditional insulin therapies (conventional therapy and multiple daily injections) and does not appear to be associated with significant adverse outcomes. Additional studies are needed to further examine the relative risks of CSII therapy, including the potential psychosocial impact of this technologically advanced therapy.
A new approach to estimate household food demand with panel data

ABSTRACT This article develops a model of food demand in which the quality and quantity of food purchased and the inter-purchase time are determined simultaneously. We use this model to explore the relationship between a household’s cereal purchases and its demographic variables. Households eligible to participate in the U.S. Government Special Supplemental Nutrition Program for Women, Infants, and Children are found to purchase a larger quantity of cereals when making a purchase and also buy more often. The model is estimated using data from Information Resources Inc.’s National Consumer Panel. These data are heavily censored at zero. The traditional approach for working with such data in demand estimation usually involves accounting for the missing unit value for non-purchase occasions and the evaluation of multivariate probabilities. However, our methodology overcomes these problems by modeling the inter-purchase time rather than modeling whether or not a purchase is made in a given time period.
Incrementally etched electrical feedthroughs for wafer-level transfer of glass lid packages

This paper reports on a simple fabrication technique to create full wafer-level transferred glass-lid encapsulations for near-hermetic packaging of MEMS devices using adhesive wafer bonding with Benzocyclobutene. Furthermore, a new technique to create low-density feedthroughs through the glass wafer for electrical interconnections from the back to the front side of the glass-lids is introduced. The through-wafer vias are fabricated by combining a mechanical etch-step by powder-blasting from the back side and a subsequent short hydrofluoric acid wet etch step. The advantage of this via technique is that the major part of the via is etched without going completely through the wafer, allowing standard surface micromachining processes on the front side of the glass wafer before the final opening of the via.
Expression of inducible nitric oxide synthase in rat experimental autoimmune myocarditis with special reference to changes in cardiac hemodynamics.

Excessive NO produced by an inducible NO synthase (iNOS) has been implicated in many types of immune-associated disorders of the cardiovascular system, but it remains to be determined whether NO plays a role in myocarditis. Thus, the significance of iNOS expression in the development of experimental autoimmune myocarditis (EAM), an animal model of human giant cell myocarditis, was investigated. Lewis rats were immunized with cardiac myosin and were killed 7, 14, 21, 28, and 49 days after immunization. The development of severe myocarditis was observed on days 14, 21, and 28 in association with significant deterioration of hemodynamics determined by cardiac catheterization, which peaked on day 21. In parallel with histological severity of myocarditis and deterioration of cardiac performance, iNOS activity in the heart measured by [14C]L-citrulline formation was markedly increased on days 14, 21, and 28. The expression of iNOS was confirmed by immunoblotting and was localized to the infiltrating inflammatory cells found in the vicinity of necrotic myocytes by immunohistochemical analysis. Aminoguanidine, a selective inhibitor of iNOS, significantly decreased the iNOS activity (1.04 +/- 0.37 compared with 29.1 +/- 8.62 pmol.min-1.mg protein-1 in untreated myosin-immunized rats, P < .01) and effectively attenuated histopathological changes of EAM on day 21. Hemodynamic parameters were also improved from 64 +/- 3 to 89 +/- 3 mm Hg for mean blood pressure, from 80 +/- 2 to 113 +/- 4 mm Hg for left ventricular systolic pressure, from 7.8 +/- 0.3 to 3.2 +/- 0.3 mm Hg for left ventricular end-diastolic pressure, from 2867 +/- 137 to 4180 +/- 102 mm Hg/s for +dP/dt, and from 2717 +/- 132 to 4180 +/- 184 mm Hg/s for -dP/dt (P < .01). The values after aminoguanidine treatment were not significantly different from the control values. These results suggest an important role for NO in mediating pathophysiological changes in myocarditis of autoimmune origin.
Process research : how multiple perspectives contribute to an organizational identity

Identity is problematic and the dynamics of identity need to be better understood. But when examining developments in organizational identity usually the why-question is asked. The how-question is underexposed in literature on the topic of organizational identity. The objective of this research is to give a comprehensible clarification of the process that precedes a develop-ment in identity. This paper explains what social mechanisms contribute to the composition of an organizational identity on the basis of a case study. The student organization of VGS-Nederland has been examined to find out from which perspec-tives identity is determined and how they contribute to it. The two perspectives that contribute to an organizational identity according to current literature have been examined to what extent they determined the identity of VGS-Nederland. But this could not explain the developments in identity fully, therefore two new perspectives have been found that contributed to the identity of VGS-Nederland. Semi-structured interviews have been conducted to create an image of determining identity in the last 25 years. The interviews and archival data resulted in these four perspectives that can be used for a better understanding of the composition of an organizational identity.
Apolipoprotein E Inhibits Serum-stimulated Cell Proliferation and Enhances Serum-independent Cell Proliferation*

Independently of its role in lipid homeostasis, apolipoprotein E (apoE) inhibits cell proliferation. We compared the effects of apoE added to media (exogenous apoE) with the effects of stably expressed apoE (endogenous apoE) on cell proliferation. Exogenous and endogenous apoE increased population doubling times by 30–50% over a period of 14 days by prolonging the G1 phase of the cell cycle. Exogenous and endogenous apoE also decreased serum-stimulated DNA synthesis by 30–50%. However, apoE did not cause cell cycle arrest; both apoE-treated and control cells achieved equivalent saturation densities at 14 days. Further analyses demonstrated that exogenous and endogenous apoE prevented activation of MAPK but not induction of c-fosexpression in response to serum growth factors. Endogenous (but not exogenous) apoE altered serum concentration-dependent effects on proliferation. Whereas control (non-apoE-expressing) cell numbers increased with increasing serum concentrations (1.6-fold for every 2-fold increase in serum), apoE-expressing cell numbers did not differ as serum levels were raised from 2.5 to 10%. In addition, in low serum (0.1%), apoE-expressing cells had elevated DNA synthesis levels compared with control cells. We conclude that apoE does not simply inhibit cell proliferation; rather, the presence of apoE alters the response to and requirement for serum mitogens.
Shipping to Heterogeneous Customers with Competing Carriers

Problem definition: We study a shipper transporting and selling a short life-cycle product to a destination market via two competing transportation service providers (i.e., carriers). The two carriers offer distinct speeds and competing freight rates, whereas customers in the destination market obtain higher utility if they receive the product earlier and their time preferences are heterogeneous. Academic/practical relevance: Perishable products are commonly shipped via multiple means of transport. The faster the mode of transport, the more expensive it is, but speed enables the product to reach the market with higher quality. In addition to the trade-off between speed and cost, the competition between carriers can also influence the shipper's transportation procurement strategies. Our model highlights the implications of carrier competition in a dual sourcing problem. Methodology: We analyze a two-stage game in which carriers first compete on freight rates, and then the shipper determines the shipping quantities. Results: We show that the shipper may benefit from product differentiation via dual-mode shipping, in which the shipment that arrives earlier is sold at a premium price. In equilibrium, the shipper's profit can be U-shaped in the speed difference between carriers. Dual sourcing may be inferior to simply restricting a single shipping service in a winner-take-all fashion. Managerial implications: This study reveals an underlying trade-off between the operational advantage from product differentiation and the cost advantage from carrier competition. To benefit from either of these advantages, a shipper should use two carriers with either very distinct or very similar speeds. Single sourcing may bring an additional cost advantage that outweighs the value of production differentiation through dual sourcing.
Inflammatory mechanisms in major depressive disorder

Purpose of review As the ‘monoamine hypothesis of depression’ fails to explain all aspects of major depression, additional causes are being investigated. Several observations suggest that inflammatory mechanisms pay a role in the cause of major depressive disorder (MDD). This article reviews their role in major depression. Recent findings Recent studies support the concept that inflammatory mechanisms play a crucial role in the pathomechanisms of major depression. Major depression shares similarities with ‘sickness behavior’, a normal response to inflammatory cytokines. Elevations in proinflammatory cytokines and other inflammation-related proteins in major depression were found in plasma and cerebrospinal fluid (CSF) as well as in postmortem studies. Elevated levels of proinflammatory cytokines persist after clinical symptoms of depression are in remission and can also predict the onset of a depressive episode. Antidepressant treatment can lead to a normalization of elevated cytokine levels in major depression. Finally, we understand how inflammatory mechanisms affect the metabolism of tryptophan and how nonsteroidal antiinflammatory drugs (NSAIDs) can interfere with the effects of antidepressants. Summary Further studies are needed to fully understand the role of inflammatory mechanisms in major depression and the potential treatment implications.
Treatment of Pain in Pregnancy

BackgroundThe treatment of pain during pregnancy other than that of labor is a clinical issue that has not been addressed in a systematic manner. Materials and MethodsTo assess current knowledge, a review of the human and animal literatures was undertaken using MEDLINE. In addition, the dynamics of three pharmacological compartments, the mother, the placenta, and the fetus, and fate of drugs given in pregnancy, was reviewed. ResultsThe literature review yielded little information except for a few case studies in which opiates, nonsteroidal anti-inflammatory drugs, antidepressants, &mgr; agonists, and anticonvulsants were used in the treatment of pain in pregnancy. In contrast, there is extensive information in the addiction medicine literature concerning the use of opioids in recovering pregnant addicts. Methadone, buprenorphine, and morphine have been used to treat women seeking recovery from opioids, and neonatal outcomes have been closely monitored with no evidence of harm to the newborn. ConclusionsExperience in women seeking recovery from opioids and their newborns illustrates that opioids are an effective and safe pharmacological option for the treatment of pain during pregnancy. Controlled studies are needed to expand knowledge in this clinical area.
SSB, Encoding a Ribosome-Associated Chaperone, Is Coordinately Regulated with Ribosomal Protein Genes

ABSTRACT Genes encoding ribosomal proteins and other components of the translational apparatus are coregulated to efficiently adjust the protein synthetic capacity of the cell. Ssb, a Saccharomyces cerevisiae Hsp70 cytosolic molecular chaperone, is associated with the ribosome-nascent chain complex. To determine whether this chaperone is coregulated with ribosomal proteins, we studied the mRNA regulation of SSB under several environmental conditions. Ssb and the ribosomal protein rpL5 mRNAs were up-regulated upon carbon upshift and down-regulated upon amino acid limitation, unlike the mRNA of another cytosolic Hsp70, Ssa. Ribosomal protein and Ssb mRNAs, like many mRNAs, are down-regulated upon a rapid temperature upshift. The mRNA reduction of several ribosomal protein genes and Ssb was delayed by the presence of an allele, EXA3-1, of the gene encoding the heat shock factor (HSF). However, upon a heat shock theEXA3-1 mutation did not significantly alter the reduction in the mRNA levels of two genes encoding proteins unrelated to the translational apparatus. Analysis of gene fusions indicated that the transcribed region, but not the promoter of SSB, is sufficient for this HSF-dependent regulation. Our studies suggest that Ssb is regulated like a core component of the ribosome and that HSF is required for proper regulation of SSB and ribosomal mRNA after a temperature upshift.
Variability of the metabolic effect of soluble insulin and the rapid-acting insulin analog insulin aspart.

OBJECTIVE To study the intra- and interindividual variability of the metabolic activity of soluble insulin and of the rapid-acting insulin analog insulin aspart after subcutaneous injection. RESEARCH DESIGN AND METHODS A total of nine healthy male volunteers received subcutaneous injections of soluble insulin (0.2 U/kg) in the abdominal region on each of the four study days. Another 10 volunteers received an injection of insulin aspart four times. Glucose infusion rates necessary to neutralize the blood glucose-lowering effect of the administered insulin were registered during euglycemic glucose clamps (blood glucose 5.0 mmol/l; basal intravenous insulin infusion 0.15 mU x kg(-1) x min(-1) over the subsequent 600 min. We investigated the variation in metabolic activity by calculating coefficients of variation (CVs). RESULTS In comparison to soluble insulin, subcutaneous injections of insulin aspart led to a more rapid onset of action and a shorter duration of action. Subcutaneous injection of the insulin preparations resulted in intraindividual CVs of the summary measures between 10 and 30% (soluble insulin vs. insulin aspart: maximal metabolic activity 15+/-7 vs. 16+/-10%, time to maximal metabolic activity 14+/-10 vs. 11+/-6%; NS between the preparations [means +/- SD]). The decline to half-maximal activity after maximal activity showed a lower intraindividual CV with insulin aspart (19+/-9 vs. 11+/-5%; P = 0.018). The interindividual CVs were higher than the intraindividual CVs (26 vs. 28, 23 vs. 19, and 26 vs. 17%). Generally, the pharmacodynamic variability was higher than the pharmacokinetic variability. For the pharmacokinetic measures, the intra- and interindividual variability in t(max) was lower for insulin aspart than for soluble insulin. CONCLUSIONS The metabolic effect of soluble insulin shows an intraindividual variability of 10-20% in healthy volunteers, even under strictly controlled experimental conditions. The overall variability of action of insulin aspart was comparable to that of soluble insulin.
IDDM in Rats Induced by Thymectomy and Irradiation

A diabetic syndrome closely resembling human IDDM has been induced in rats of specific pathogen-free origin by a combination of thymectomy and irradiation, with an overall incidence (10 wk postirradiation) in female rats of 34% for acute disease and 47% for islet lesions. Males were slightly more susceptible than females. Clinical features of the syndrome included hyperglycemia, insulinopenia, ketosis, and lipidemia, and corresponding islet pathology ranged from diffuse atrophy to focal atrophy and insulitis. Onset was usually acute, and the disease fatal unless early insulin therapy was initiated. Lymphocytic thyroiditis also was observed in a proportion of thymectomized and irradiated rats (49% in females) over the same period but with no apparent correlation to the occurrence of diabetes. A significant decrease in the incidence of disease was found in thymectomized and irradiated rats of conventional origin when compared with genetically identical specific pathogen-free rats, implicating a role for environmental factors in disease susceptibility. Diabetes inducement also was found to be strain related but not RT1u dependent. Both clinical signs and islet lesions were inhibited by early reconstitution of thymectomized and irradiated animals with syngeneic lymphoid cells from normal donors. Islet lesions and glucose intolerance could be transferred to syngeneic recipients by concanavalin A–activated lymphoid cells from acute diabetic donors. The close similarities between this experimental syndrome induced by immunological manipulation and the clinical condition in humans provide further evidence for an immune-mediated pathogenesis for IDDM.