Gamma knife surgery for dural arteriovenous shunts: 25 years of experience.
The aim of this study was to assess the clinical efficacy of gamma knife surgery (GKS) in the treatment of dural arteriovenous shunts (DAVSs). From a database of more than 1600 patients with intracranial arteriovenous shunts that had been treated with GKS, the authors retrospectively and prospectively identified 53 patients with 58 DAVSs from the period between 1978 and 2003. Four patients were lost to follow-up evaluation and were excluded from the series. Thus, this study is based on the remaining 49 patients with 52 DAVSs. Thirty-six of the shunts drained into the cortical venous system, either directly or indirectly, and 22 of these were associated with intracranial hemorrhage on patient presentation. The mean prescription radiation dose was 22 Gy (range 10-28 Gy). All patients underwent a clinical follow-up examination. In 41 cases of DAVS a follow-up angiography study was performed. At the 2-year follow-up visit, 28 cases (68%) had angiographically proven obliteration of the shunt and in another 10 cases (24%) there was significant flow regression. Three shunts remained unchanged. There was one immediate minor complication related to the administration of radiation. Furthermore, one patient had a radiation-induced complication 10 years after treatment, although she recovered completely. There was one posterior fossa bleed 2 months after radiosurgery; a hematoma, as well as a lesion, was evacuated, and the patient recovered uneventfully. A second patient had an asymptomatic occipital hemorrhage approximately 6 months postradiosurgery. The clinical outcome after GKS was significantly better than that in patients with naturally progressing shunts (p < 0.01, chi-square test); figures on the latter have been reported previously. Gamma knife surgery is an effective treatment for DAVSs, with a low risk of complications. Major disadvantages of this therapy include the time elapsed before obliteration and the possibility that not all shunts will be obliterated. Cortical venous drainage from a DAVS, a risk factor for intracranial hemorrhage, is therefore a relative contraindication. Consequently, GKS can be used in the treatment of both benign DAVSs with subjectively intolerable bruit and aggressive DAVSs not responsive to endovascular treatment or surgery.


Use of standardised patients to assess quality of healthcare in Nairobi, Kenya: a pilot, cross-sectional study with international comparisons.
The quality of clinical care can be reliably measured in multiple settings using standardised patients (SPs), but this methodology has not been extensively used in Sub-Saharan Africa. This study validates the use of SPs for a variety of tracer conditions in Nairobi, Kenya, and provides new results on the quality of care in sampled primary care clinics. We deployed 14 SPs in private and public clinics presenting either asthma, child diarrhoea, tuberculosis or unstable angina. Case management guidelines and checklists were jointly developed with the Ministry of Health. We validated the SP method based on the ability of SPs to avoid detection or dangerous situations, without imposing a substantial time burden on providers. We also evaluated the sensitivity of quality measures to SP characteristics. We assessed quality of practice through adherence to guidelines and checklists for the entire sample, stratified by case and stratified by sector, and in comparison with previously published results from urban India, rural India and rural China. Across 166 interactions in 42 facilities, detection rates and exposure to unsafe conditions were both zero. There were no detected outcome correlations with SP characteristics that would bias the results. Across all four conditions, 53% of SPs were correctly managed with wide variation across tracer conditions. SPs paid 76% less in public clinics, but proportions of correct management were similar to private clinics for three conditions and higher for the fourth. Kenyan outcomes compared favourably with India and China in all but the angina case. The SP method is safe and effective in the urban Kenyan setting for the assessment of clinical practice. The pilot results suggest that public providers in this setting provide similar rates of correct management to private providers at significantly lower out-of-pocket costs for patients. However, comparisons across countries are sensitive to the tracer condition considered.


Immunomodulatory therapies for cystic fibrosis.
Progressive pulmonary disease is the primary cause of morbidity and mortality in adult patients with cystic fibrosis (CF). The decrease in lung function associated with infection with Pseudomonas aeruginosa has been related to the severity of pulmonary inflammation. Thus therapies that reduce pulmonary inflammation may prove to be clinically efficacious. Therapeutic interventions that target pulmonary inflammation may be directed either at the infecting organism, especially P aeruginosa, or at host responses. Eradication of P aeruginosa from the airways of patients with CF has not been accomplished. However, reduction of the burden of P aeruginosa or modification of virulence factors are practical goals. Normalization of the host response ultimately depends on correction of the molecular defect. Until then, therapies are being investigated that may modulate pulmonary inflammation. These include therapies aimed at compensating for the defect in ion transport, down-regulating inflammatory cell responses, inhibiting host inflammatory products, or altering airway secretions. Preliminary data suggest that each of these approaches may have clinical efficacy. Large, multicenter trials addressing these issues are presently ongoing and hold the promise for continued improvement in the clinical course of patients with CF.


Brain Donation by Proxy: Are There Predictors in Neurodegenerative Dementia?
To describe the frequency and predictors of brain donation by relatives in patients with neurodegenerative dementia. Database review and quantitative analysis. The Alzheimer Center Reina Sofia Foundation (ACRSF), a center devoted to the care and research of patients with neurodegenerative dementia. Patients with signed consent for participation in the ACRSF research program. A set of 38 demographic, clinical, and social variables related to patient and closest relative, which were collected by the ACRSF multidisciplinary team upon patient admission. Admission data were available for 198 patients who entered the ACRSF research program; 85 of them (42.9%) died during follow-up. Mean age (SD) at admission was 82.3 (6.8) years and 80.8% of the patients were female. Family link between patient and closest relative was spouse or partner (12.0%), son or daughter (74.9%), or other link (13.1%). Brain was obtained from 56 patients (65.9%). Consent by legal representative and patient's depressive symptoms were more frequent in the donors (p<0.05, corrected) and trend was observed for more aberrant motor symptoms in the donors (p<0.05, uncorrected). A high rate of brain donation was achieved, probably due to the unique characteristics of the ACRSF and consent for research policy. Wish of alleviating suffering, as well as general interest in dementia research, possibly exerted an influence in brain donation. More research is needed to ascertain the values, motivations, and circumstances that may lead to brain donation by proxy in neurodegenerative dementia.


Patch clamping the outer mitochondrial membrane.
Intact giant mitochondria isolated from the liver of mice fed a diet containing cuprizone were studied using patch microelectrodes. The current-voltage curves were nonlinear, suggesting the presence of voltage-sensitive channels. In the negative range of voltage, the channels appear to close with increasing magnitude of the voltage. The dependence of the conductance on voltage is similar to that of the outer membrane channels (VDAC) studied in planar bilayers. Occasionally, over a narrow range of positive potentials, the conductance also decreases as in the bilayer studies. However, more frequently the conductance increases sharply in a completely reversible manner at potentials greater than 10 to 20 mV. The increase in conductance with voltage may be interpreted as a major rearrangement of membrane components. Qualitatively comparable results were obtained using fused outer membranes isolated from Neurospora mitochondria. The behavior of VDAC is affected by treatment with succinic anhydride or the polyanion, polymethacrylate, maleate, styrene (1:2:3). We have found similar effects in the negative range of potentials in patches from giant mitochondria treated in the same fashion.


[NMDA receptor and long-term potentiation].
In order to evaluate the role of the NMDA receptor redox site in long-term potentiation (LTP), we have investigated the effects of two redox reagents, 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) and tris(carboxyethyl)phosphine (TCEP) on the induction and expression of various forms of LTP. DTNB a thiol-oxidizing agent, irreversibly reduces by 50% NMDA receptor EPSP. In the presence of DTNB, the induction of tetanic and anoxic LTP are prevented. When tetanic or anoxic LTP were generated first, DTNB completely reverses the potentiation and TCEP a disulfide-reducing agent restores LTP to its initial level. These redox agents have no effect on AMPA synaptic transmission and did not significantly modify the induction and the expression of tetanic AMPA-LTP. These results suggest that thiol-oxidizing compounds might be useful for the treatment of cerebral ischemia.


The Interstitial Pathways as the Substrate of Consciousness: A New Synthesis.
This paper considers three classes of analyses of the nature of consciousness: abstract theories of the functional organization of consciousness, and concrete proposals as to the neural substrate of consciousness, while providing a rationale for contesting non-neural and transcendental conceptualizations of consciousness. It indicates that abstract theories of the dynamic core of consciousness have no force unless they are grounded in the physiology of the brain, since the organization of dynamic systems, such as the Sun, could equally well qualify as conscious under such theories. In reviewing the wealth of studies of human consciousness since the mid-20th century, it concludes that many proposals for the particular neural substrate of consciousness are insufficient in various respects, but that the results can be integrated into a novel scheme that consciousness extends through a subcortical network of interlaminar structures from the brainstem to the claustrum. This interstitial structure has both the specificity and the extended connectivity to account for the array of reportable conscious experiences.


Isolation of partly heme devoid but not fully carbon monoxide reacting cytochrome b, from beef heart mitochondria.
An easy and rapid preparation of cytochrome b from beef heart mitochondria is described. The method is based on four fractionation steps in the presence of bile acid and one step DE chromatography. The isolated cytochrome shows only partial reactivity with carbon monoxide; up to a level of 35 micrograms protein applied, it moves as a single band on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, with an apparent molecular weight of 32000 +/- 1000. A ratio of 14.1 +/- 0.8 nmol heme b/mg protein has been found for isolated cytochrome, indicating that good part of heme b is lost during the isolation. The low number of steps required for the isolation could allow the identification of the stage(s) responsible for the loss of heme moiety and indicate preparation improvements.


Importing ArrayExpress datasets into R/Bioconductor.
ArrayExpress is one of the largest public repositories of microarray datasets. R/Bioconductor provides a comprehensive suite of microarray analysis and integrative bioinformatics software. However, easy ways for importing datasets from ArrayExpress into R/Bioconductor have been lacking. Here, we present such a tool that is suitable for both interactive and automated use. The ArrayExpress package is available from the Bioconductor project at http://www.bioconductor.org. A users guide and examples are provided with the package.


Elucidating the mechanism of the surface functionalization dependent neurotoxicity of graphene family nanomaterials.
Graphene family nanomaterials (GFNs) have shown great potential for biological and environmental applications; however, their future use has been debated due to their reported potential neurotoxicity. Moreover, the effects of surface functionalization on their biological end points are largely unknown. Here, we compared the effects of reduced graphene oxide (RGO), and carboxylated (G-COOH), hydroxylated (G-OH) and aminated (G-NH2) graphene nanosheets on human neuroblastoma cells (SK-N-SH). All GFNs inhibited cellular growth at concentrations of 0.1-10 mg L-1 after 24 h exposure. The toxicity was attenuated over longer exposure times, with the exception of G-NH2. Although the overall acute toxicity followed the order: G-OH ≈ G-COOH > RGO > G-NH2, G-NH2 induced more persistent toxicity and more metabolic disturbance compared to the other GFNs, with lipid and carbohydrate metabolism being the most affected. The potential for physical disruption of the lipid membrane and oxidative damage induced by GFNs varied with different functionalization, which accounts for the observed differences in neurotoxicity. This study provides significant insights into the neurological effects of GFNs, and suggests that G-NH2 is not as safe as reported in many previous studies. The neurological effect of GFNs over longer term exposure should be considered in future studies.


Mutation accumulation and developmental lineages in normal and Down syndrome human fetal haematopoiesis.
Children show a higher incidence of leukemia compared to young adolescents, yet their cells have less age-related (oncogenic) somatic mutations. Newborns with Down syndrome have an even higher risk of developing leukemia, which is thought to be driven by mutations that accumulate during fetal development. To characterize mutation accumulation in individual stem and progenitor cells of Down syndrome and karyotypically normal fetuses, we clonally expanded single cells and performed whole-genome sequencing. We found a higher mutation rate in haematopoietic stem and progenitor cells during fetal development compared to the post-infant rate. In fetal trisomy 21 cells the number of somatic mutations is even further increased, which was already apparent during the first cell divisions of embryogenesis before gastrulation. The number and types of mutations in fetal trisomy 21 haematopoietic stem and progenitor cells were similar to those in Down syndrome-associated myeloid preleukemia and could be attributed to mutational processes that were active during normal fetal haematopoiesis. Finally, we found that the contribution of early embryonic cells to human fetal tissues can vary considerably between individuals. The increased mutation rates found in this study, may contribute to the increased risk of leukemia early during life and the higher incidence of leukemia in Down syndrome.


Initial assessment of the role of CXC chemokine receptor 4 after polytrauma.
CXC chemokine receptor (CXCR)-4 agonists have been shown to attenuate inflammation and organ injury in various disease models, including trauma/hemorrhage. The pathophysiological role of CXCR4 during the early response to tissue injury, however, remains unknown. Therefore, we investigated the effects of AMD3100, a drug that antagonizes binding of stromal cell-derived factor (SDF)-1α and ubiquitin to CXCR4 during the initial response to polytrauma in pigs. Fifteen minutes before polytrauma (femur fractures/lung contusion; control: sham), 350 nmol/kg AMD3100, equimolar AMD3100 and ubiquitin (350 nmol/kg each) or vehicle were administered intravenously. After a 60-min shock period, fluid resuscitation was performed for 360 min. Ubiquitin binding to peripheral blood mononuclear cells was significantly reduced after intravenous AMD3100. SDF-1α plasma levels increased transiently >10-fold with AMD3100 in all animals. In injured animals, AMD3100 increased fluid requirements to maintain hemodynamics and enhanced increases in peripheral blood granulocytes, lymphocytes and monocytes, compared with its effects in uninjured animals. Cytokine release from leukocytes in response to Toll-like receptor (TLR)-2 and TLR-4 activation was increased after in vitro AMD3100 treatment of normal whole blood and after in vivo AMD3100 administration in animals subjected to polytrauma. Coadministration of AMD3100/ubiquitin reduced lactate levels, prevented AMD3100-induced increases in fluid requirements and sensitization of the tumor necrosis factor (TNF)-α and interleukin (IL)-6 release upon TLR-2/4 activation, but did not attenuate increases in leukocyte counts and SDF-1α plasma levels. Our findings suggest that CXCR4 controls leukocyte mobilization after trauma, regulates leukocyte reactivity toward inflammatory stimuli and mediates protective effects during the early phase of trauma-induced inflammation.


Frequency division multiplexing readout of a transition edge sensor bolometer array with microstrip-type electrical bias lines.
We demonstrate multiplexed readout of 43 transition edge sensor (TES) bolometers operating at 90 mK using a frequency division multiplexing (FDM) readout chain with bias frequencies ranging from 1 to 3.5 MHz and a typical frequency spacing of 32 kHz. We improve the previously reported performance of our FDM system by two important steps. First, we replace the coplanar wires with microstrip wires, which minimize the cross talk from mutual inductance. From the measured electrical cross talk (ECT) map, the ECT of all pixels is carrier leakage dominated. Only five pixels show an ECT level higher than 1%. Second, we reduce the thermal response speed of the TES detectors by a factor of 20 by increasing the heat capacity of the TES, which allows us to bias all TES detectors below 50% in transition without oscillations. We compare the current-voltage curves and noise spectra of the TESs measured in single-pixel mode and multiplexing mode. We also compare the noise equivalent power (NEP) and the saturation power of the bolometers in both modes, where 38 pixels show less than 10% difference in NEP and 5% difference in saturation power when measured in the two different modes. The measured noise spectrum is in good agreement with the simulated noise based on measured parameters from an impedance measurement, confirming that our TES is dominated by phonon noise.


Chromatic patterns of cone photoreceptors. 1976 Glenn A. Fry Award Lecture.
The operations of vertebrate visual neurons must eventually be expressed in terms of the spectral classes and spatial distributions of photoreceptors that constitute inputs to those neurons. By using a cytochemical probe that permits visualization of the responses of cone photoreceptors to light stimulation, it has been possible to describe the spectral classes and spatial patterns of cones in a lower vertebrate, the goldfish, and in a primate, the baboon. This information is an important first step in analyzing the connections between cones and retinal neurons.


THE ELIMINATION OF ADMINISTERED ZINC IN PANCREATIC JUICE, DUODENAL JUICE, AND BILE OF THE DOG AS MEASURED BY ITS RADIOACTIVE ISOTOPE (Zn).
The excretion of intravenously injected labeled zinc into pancreatic juice, duodenal juice, and bile of the dog was investigated. The use of the radioactive isotope permitted the injection of minute amounts of zinc, amounts that were negligible when compared with the total amount of zinc already present in the animal. 1. A large proportion of the injected Zn(65) was eliminated by way of the external secretion of the pancreas. As much as 11 per cent was excreted in the pancreatic juice in 14 days. 2. Very little of the administered radiozinc appeared in the bile. The maximum excretion observed for this pathway was 0.4 per cent in 8 days. 3. Radiozinc was also found in large amounts in the juice obtained from an isolated loop of the duodenum.


Core-shell PdAu nanocluster catalysts to suppress sulfur poisoning.
Reducing sulfur poisoning is significant for maintaining the catalytic efficiency and durability of heterogeneous catalysts. We screened PdAu nanoclusters with specific Pd : Au ratios based on Monte Carlo simulations and then carried out density functional calculations to reveal how to reduce sulfur poisoning via alloying. Among various nanoclusters, the core-shell structure Pd13Au42 (Pd@Au) exhibits a low adsorption energy of SO2 (-0.67 eV), comparable with O2 (-0.45 eV) and lower than CO (-1.25 eV), thus avoiding sulfur poisoning during the CO catalytic oxidation. Fundamentally, the weak adsorption of SO2 originates from the negative d-band center of the shell and delocalized charge distribution near the Fermi level, due to the appropriate charge transfer from the core to shell. Core-shell nanoclusters with a different core (Ni, Cu, Ag, Pt) and a Pd@Au slab model were further constructed to validate and extend the results. These findings provide insights into designing core-shell catalysts to suppress sulfur poisoning while optimizing catalytic behaviors.


Evaluating quality of life in hypertensive patients.
The treatment of arterial hypertension is symptomatic in 90-95% of patients, and therefore it must be administered throughout life. At the beginning of pharmacologic treatment when only patients with severe or malignant hypertension are treated, the goal is almost exclusively limited to blood pressure reduction. Thereafter, when the treatment is extended to patients with mild and moderate hypertension, other aims in addition to blood pressure reduction, are evaluated and among these is the impact of pharmacologic blood pressure lowering on the quality of life. The quality of life is recognized as a multi-factorial variable and can be subdivided into six domains. The methodology used to evaluate the quality of life should use valid, repeatable, and sensitive tools. A metaanalysis of well selected and comparable trials has shown that antihypertensive treatment, as a whole, has a small but positive impact on many domains of the quality of life. Furthermore, it appears that converting-enzyme inhibitors, beta-blockers, calcium antagonists, and diuretics cause a statistically significant improvement of quality of life, while centrally acting alpha 1-agonists and direct vasodilators show only a positive trend. Although the comparison among two or more drugs with regard to quality of life is more difficult, it appears from a personal review that converting-enzyme inhibitors and calcium antagonists cause a greater improvement. These two classes of antihypertensive agents have been shown to improve the quality of life in elderly hypertensive patients, together with significant blood pressure reduction. Finally, the incidence of drop-outs and side effects cannot be considered a valid means of evaluation of the quality of life.


Upside-down stomach in paraesophageal hernia: A case report.
Paraesophageal hernias, accounting for a mere 5% to 10% of all hiatal hernias, occasionally present an exceedingly uncommon yet gravely consequential complication characterized by the inversion of the stomach. Delving into the clinical manifestations and optimal therapeutic approaches for patients afflicted by this condition merits substantial exploration. A 60-year-old man was referred to our hospital with acute onset of severe epigastric pain, abdominal distension, and vomiting. A chest radiograph unveiled an elevated left diaphragmatic dome accompanied by a pronounced rightward shift of the mediastinum. Subsequent abdominal computed tomography imaging delineated the migration of the stomach, spleen, and colon into the left hemithorax, facilitated by a significant diaphragmatic defect. The diagnosis of a giant paraesophageal hernia with complete gastric inversion was established through a comprehensive evaluation of the patient's clinical manifestations and imaging findings. Surgical intervention was performed on the patient. During the procedure, a left diaphragmatic defect measuring approximately 10 × 8 cm was identified and meticulously repositioned, followed by the repair of the diaphragmatic hernia. The herniated contents comprised the pancreas, stomach, spleen, a segment of the colon, and a portion of the greater omentum. The patient experienced a smooth postoperative recuperation and was discharged 12 days following the surgical procedure. Subsequently, during a 7-month follow-up period, the patient continued to exhibit favorable progress and recovery. Paraesophageal hernias are rare, and the presence of an inverted stomach in a giant paraesophageal hernia is exceptionally uncommon. Clinical presentation lacks distinct features and can lead to misdiagnosis. This case emphasizes the importance of timely surgical intervention guided by imaging, offering valuable clinical insights.


Degenerate MHC restriction reveals the contribution of class I MHC molecules in determining the fine specificity of CTL recognition of an immunodominant determinant of HIV-1 gp160 V3 loop.
The novel allogeneic presentation of an immunodominant determinant within the HIV-1 gp160 V3 loop by three different class I MHC molecules to the same CD8+ CTL is used to study the influence of the MHC molecule on the fine specificity of CTL recognition. We previously reported that four distinct class I molecules of H-2d,u,p,q presented the V3 decapeptide P18-I10 (RGPGRAFVTI) to CTL. Surprisingly, we found that H-2d,u,p cells mutually cross-present the P18-I10 peptide to allogeneic CTL clones of each of the other haplotypes, whereas none of these cross-presents to H-2q CTL, nor do H-2q targets present to CTL of the other haplotypes. Here, we explore the critical amino acid residues for the cross-presentation using 10 variant peptides with single amino acid substitutions. The fine specificity examined using these mutant peptides presented by the same MHC class I molecule showed striking similarity among the CTL of each haplotype, expressing either V beta 8.1 or V beta 14. In contrast, the fine specificity is different between the distinct MHC class I molecules even for the lysis by the same CTL, as shown by reciprocal effects of the same substitutions. Thus, peptide fine specificity of a single TCR is influenced by changes in the class I MHC molecules presenting the Ag.


Cryptococcal pleuritis with pleural effusion as the only clinical presentation in a patient with hepatic cirrhosis: A case report and literature review.
Cryptococcosis is a significant life-threatening fungal infection in worldwide, mainly reported in immunocompromised patients. Pleural effusion presentation of cryptococcal infection as the only clinical presentation is rarely seen in pulmonary cryptococcosis, which may lead to be misdiagnosed, and the study on this subject will provide further insights. A 64-year-old man was hospitalized in our department and diagnosed as hepatic B cirrhosis. A computed tomography (CT) of the thorax showed a massive right pleural effusion without pulmonary parenchymal abnormalities. He was started on empirical treatment for pleural tuberculosis (TB). However, during his hospitalization, a right pleural effusion developed and fever was not controlled. On day 14 admission, pleural fluid cultured positive for Cryptococcus neoformans. The C neoformans isolate belonged to ST5 and molecular type VNI (var. grubii). The patient was diagnosed with cryptococcal pleuritis, then amphotericin B and fluconazole were administrated. Finally, the patient was improved and discharged from our hospital. Similar cases in cryptococcal pleuritis patients with pleural effusion as the only clinical presentation in the literature are also reviewed. Through literature review, we recommend that pleural effusion cryptococcal antigen test should be used to diagnose cryptococcal pleuritis to reduce misdiagnosis. The early administration of antifungal drug with activity to Cryptococcus seemed beneficial in preventing dissemination of cryptococcosis.


Alkynylcyclohexanol chairs and twist-boats: Co(2)(CO)(6) as a conformational switch.
Treatment of 1-[axial]-(trimethylsilylethynyl)cyclohexan-1-ol with dicobalt octacarbonyl results in a conformational ring flip such that the bulky dicobalt-alkyne cluster moiety now occupies the favored equatorial site. However, when a 4-tert-butyl substituent is present, the coordinated alkynyl group retains its original axial or equatorial position. Complexation of trans-[diaxial]-1,4-bis(triphenylsilylethynyl)cyclohexane-1,4-diol brings about a chair-to-chair conformational inversion such that both cluster fragments now occupy equatorial sites. In contrast, cis-1,4-bis(triphenylsilylethynyl)cyclohexane-1,4-diol reacts with Co(2)(CO)(8) to yield the twist-boat conformer in which the two axial hydroxy substituents exhibit intra-molecular hydrogen bonding. Likewise, the corresponding reaction of cis-1,4-bis(trimethylsilylethynyl)cyclohexane-1,4-diol with Co(2)(CO)(8) leads to a twist-boat, but in this case, the molecules are linked through inter-molecular hydrogen bonds. Eight of these cobalt clusters have been characterized by X-ray crystallography, and the potential use of twist-boats in synthesis is discussed.


Efficacy and safety of glucokinase activators for type 2 diabetes mellitus therapy: a meta-analysis of double-blind randomized controlled trials.
We aimed to assess the efficacy and safety of oral glucokinase activator (GKA) in the treatment of type 2 diabetes mellitus (T2DM). We searched and collected randomized controlled trials (RCTs) of glucokinase activators in the treatment of T2DM from PubMed, ClinicalTrails, Cochrane Library, Web of Science, and CNKI databases. Revman5.3 software was used to do the meta-analysis, and the Cochrane tool was used to evaluate the risk of bias in the included RCTs. Seven double-blind RTCs were included in the final analysis, with a total of 762 patients. For the efficacy, the meta-analysis found that GKAs lowed the levels of fasting blood glucose (FPG) (mean difference -0.71, 95% CI: -1.11 to -0.31, based on 459 patients from 5 works of literature) and glycated hemoglobin (HbA1c) (mean difference: -0.65%, 95% CI: -0.82 to -0.48, based on 570 patients from 4 works of literature). Subgroup analysis showed GKAs combined with metformin, but not used alone, reduced the levels of FPG. In terms of safety, GKAs did not affect the total rate of adverse events (AEs). GKAs did not affected the risks of diarrhea (RR 1.59, 95% CI: 0.7 to 3.65, p =0.26), headache (RR 0.96, 95% CI: 0.41-2.21, p =0.60), and nausea (RR 2.23, 95% CI: 0.55-9.12, p =0.24), but they increased the risk of hypoglycemia (RR 1.81, 95% CI: 1.35 to 2.42, p <0.0001, based on 570 patients from 4 literatures). Oral GKAs combined with metformin has an evident hypoglycemic effect on T2DM, and they seem to be relatively good tolerant. Further clinical studies are still necessary to explore its long-term efficacy and safety.


X-ray imaging of differential vascular density in MMP-9-/-, PAR-1-/+, hyperhomocysteinemic (CBS-/+) and diabetic (Ins2-/+) mice.
Although protease activated receptor-1 (PAR-1) and matrix metalloproteinase-9 (MMP-9) play significant role in vascular remodelling in hyperhomocysteinemia (HHcy due to cystathionine beta synthase deficiency, CBS-/+) and diabetes, mechanism remains nebulous. We hypothesized that differential vascular density and remodelling in different vascular beds in HHcy and diabetes were responsible for an adaptive metabolic homeostasis during the pathogenesis. To test this hypothesis, vascular density in mice lacking PAR-1, MMP-9, CBS and Insulin-2 gene mutant (Ins2-/+, Akita) was measured and compared with wild type (WT, C57BL/6J) mice. The vascular density was detected by x-ray angiography using KODAK 4000 MM image station, using barium sulphate as contrasting agent. The % vascular density in the hearts of WT, CBS-/+ (HHcy), MMP-9-/-, PAR-1-/+ and Ins2-/+ (type-1 diabetes) was 100 ± 2.8, 85 ± 3.3, 90 ± 3.3, 95 ± 3.8 and 73 ± 1.7, respectively. The vascular density in CBS-/+ and Akita hearts decreased while it was increased in lungs of CBS-/+ and MMP-9-/-.There was decreased vascular density in liver and kidney of Akita mice. Vascular density in brain, kidney and mesentery was decreased in CBS-/+ mice. These findings support the notation that metabolic derangement in diabetes and HHcy causes the chronic decline and/or rarefaction in vascular density.


Hyperadrenocorticism associated with sex steroid excess.
Diagnosis of sex steroid excess or hyperadrenocorticism in dogs may be challenging. Unlike Cushing's disease, sex steroid excess may have a multitude of manifestations that differ from standard hyperadrenocorticism. In particular, the clinical scenario of a dog with sex steroid imbalance involves one of three systems: dermatologic, reproductive, or hepatic. The history of a dog with hyperadrenocorticism manifesting as sex steroid imbalance often lacks the classical clinical signs of polydipsia and polyuria. Dogs with sex steroid imbalance will often be of specific breeds such as miniature poodles and exhibit trunkal hair loss as the only sign. There is often involvement of the reproductive system, manifested as the growth of perianal adenomas in neutered male or female dogs. The most common laboratory findings consist of elevations in serum alkaline phosphatase and serum alanine transferase. The following article reviews the etiology, common signalment, clinical signs, and laboratory findings associated with atypical hyperadrenocorticism caused by sex steroid imbalance and then explores the medical, surgical, and radiation treatment options.


Conserved patterns of incomplete reporting in pre-vaccine era childhood diseases.
Incomplete observation is an important yet often neglected feature of observational ecological timeseries. In particular, observational case report timeseries of childhood diseases have played an important role in the formulation of mechanistic dynamical models of populations and metapopulations. Yet to our knowledge, no comprehensive study of childhood disease reporting probabilities (commonly referred to as reporting rates) has been conducted to date. Here, we provide a detailed analysis of measles and whooping cough reporting probabilities in pre-vaccine United States cities and states, as well as measles in cities of England and Wales. Overall, we find the variability between locations and diseases greatly exceeds that between methods or time periods. We demonstrate a strong relationship within location between diseases and within disease between geographical areas. In addition, we find that demographic covariates such as ethnic composition and school attendance explain a non-trivial proportion of reporting probability variation. Overall, our findings show that disease reporting is both variable and non-random and that completeness of reporting is influenced by disease identity, geography and socioeconomic factors. We suggest that variations in incomplete observation can be accounted for and that doing so can reveal ecologically important features that are otherwise obscured.


Mortality among US commercial pilots and navigators.
The airline industry may be an occupational setting with specific health risks. Two environmental agents to which flight crews are known to be exposed are cosmic radiation and magnetic fields generated by the aircraft's electrical system. Other factors to be considered are circadian disruption and conditions specific to air travel, such as noise, vibration, mild hypoxia, reduced atmospheric pressure, low humidity, and air quality. This study investigated mortality among US commercial pilots and navigators, using proportional mortality ratios for cancer and noncancer end points. Proportional cancer mortality ratios and mortality odds ratios were also calculated for comparison to the proportional mortality ratios for cancer causes of death. Results indicated that US pilots and navigators have experienced significantly increased mortality due to cancer of the kidney and renal pelvis, motor neuron disease, and external causes. In addition, increased mortality due to prostate cancer, brain cancer, colon cancer, and cancer of the lip, buccal cavity, and pharynx was suggested. Mortality was significantly decreased for 11 causes. To determine if these health outcomes are related to occupational exposures, it will be necessary to quantify each exposure separately, to study the potential synergy of effects, and to couple this information with disease data on an individual basis.


Potassium Deficiency-induced Changes in Stomatal Behavior, Leaf Water Potentials, and Root System Permeability in Beta vulgaris L.
Studies of the water relations of potassium deficient sugarbeet plants (Beta vulgaris L.) revealed two factors for stomatal closure. One component of stomatal closure was reversible by floating leaf discs on distilled water to relieve the water deficit in the leaves; the other component was reversible in the light by floating the leaf discs on KCl solution for 1 hour or more. Potassium-activated stomatal opening in the light was observed when the guard cells were surrounded by their normal environment of epidermal and mesophyll cells, just as observed by previous workers for epidermal strips. Leaf water potentials, like stomatal apertures, appear to be strongly related to leaf potassium concentration. Potassium-deficient plants have a greatly decreased root permeability to water, and the implications of this effect on stomatal aperture and leaf water potential are discussed. In contrast, petiole permeability to water is unaffected by potassium treatment.


Development of respiratory function in the American locust Schistocerca americana. I. Across-instar effects.
We tested the hypothesis that oxygen delivery from the atmosphere to the tissues becomes more difficult as grasshoppers increase in body size throughout development due to increases in tracheal length. If this is true, then older, larger grasshoppers should have smaller safety margins [higher critical oxygen partial pressures (P(O2)s)] for oxygen delivery than younger, smaller grasshoppers. We exposed grasshoppers of first, third and fifth instars and adults to decreasing levels of atmospheric O2 and measured their ventilatory responses. Contrary to our prediction, we found that larger grasshoppers had critical P(O2)s eight times lower than juveniles due in part to their threefold lower mass-specific metabolic rates and their ability to quadruple convective gas exchange. Adults more than doubled abdominal pumping frequency and increased tidal volume by 25% as P(O2) decreased fourfold, whereas the youngest juveniles showed no such responses. This study indicates that juveniles may be more susceptible to hypoxia in natural situations, such as exposure to high altitude or restricted burrows. Also, larger size is not necessarily correlated with a smaller safety margin for oxygen delivery in insects.


Generic patient-reported outcomes in child health research: a review of conceptual content using World Health Organization definitions.
Our aims were to (1) describe the conceptual basis of popular generic instruments according to World Health Organization (WHO) definitions of functioning, disability, and health (FDH), and quality of life (QOL) with health-related quality of life (HRQOL) as a subcomponent of QOL; (2) map the instruments to the International Classification of Functioning, Disability and Health (ICF); and (3) provide information on how the analyzed instruments were used in the literature. This should enable users to make valid choices about which instruments have the desired content for a specific context or purpose. Child health-based literature over a 5-year period was reviewed to find research employing health status and QOL/HRQOL instruments. WHO definitions of FDH and QOL were applied to each item of the 15 most used instruments to differentiate measures of FDH and QOL/HRQOL. The ICF was used to describe the health and health-related content (if any) in those instruments. Additional aspects of instrument use were extracted from these articles. Many instruments that were used to measure QOL/HRQOL did not reflect WHO definitions of QOL. The ICF domains within instruments were highly variable with respect to whether body functions, activities and participation, or environment were emphasized. There is inconsistency among researchers about how to measure HRQOL and QOL. Moreover, when an ICF content analysis is applied, there is variability among instruments in the health components included and emphasized. Reviewing content is important for matching instruments to their intended purpose.


Forecasting sudden changes in environmental pollution patterns.
The lack of reliable forecasts for the spread of oceanic and atmospheric contamination hinders the effective protection of the ecosystem, society, and the economy from the fallouts of environmental disasters. The consequences can be dire, as evidenced by the Deepwater Horizon oil spill in the Gulf of Mexico in 2010. We present a methodology to predict major short-term changes in environmental contamination patterns, such as oil spills in the ocean and ash clouds in the atmosphere. Our approach is based on new mathematical results on the objective (frame-independent) identification of key material surfaces that drive tracer mixing in unsteady, finite-time flow data. Some of these material surfaces, known as Lagrangian coherent structures (LCSs), turn out to admit highly attracting cores that lead to inevitable material instabilities even under future uncertainties or unexpected perturbations to the observed flow. These LCS cores have the potential to forecast imminent shape changes in the contamination pattern, even before the instability builds up and brings large masses of water or air into motion. Exploiting this potential, the LCS-core analysis developed here provides a model-independent forecasting scheme that relies only on already observed or validated flow velocities at the time the prediction is made. We use this methodology to obtain high-precision forecasts of two major instabilities that occurred in the shape of the Deepwater Horizon oil spill. This is achieved using simulated surface currents preceding the prediction times and assuming that the oil behaves as a passive tracer.


Somatostatin-immunoreactive nerve cell bodies and fibers in the medulla oblongata et spinalis.
Complete serial sectioning of the medulla oblongata in monkey, cat, guinea pig, and japanese dancing mouse and incubation for somatostatin-immunoreaction was carried out. Numerous regions of the medulla oblongata such as the nucleus reticularis gigantocellularis, nucleus cuneatus et gracillis, nucleus raphe magnus, nucleus tractus solitarius, nucleus vestibularis, and parts of the oliva contain dense networks of somatostatin-immunoreactive nerve fibers. Cell bodies were seen in the nucleus reticularis medullae oblongatae. In the spinal cord the sections from each segment were analyzed, showing the highest concentrations of somatostatinergic fibers in the substantia gelantinosa of the columna dorsalis. Cell bodies were seen in the zona intermedia centralis, especially in the upper cervical segments. Many positive fibers were also seen in the entire zona intermedia and the columna ventralis. Especially prominent was the immunoreactivity in the zona intermediolateralis of the thoracic segments and the columna ventralis of the lower lumbar and sacral segments.


Circulation of group 2 coronaviruses in a bat species common to urban areas in Western Europe.
Fecal samples of 211 bats representing 13 different bat species from 31 locations in the Netherlands were analyzed for the presence of coronaviruses (CoV) using a genus-wide reverse transcription (RT)-polymerase chain reaction. CoVs are known for their high potential for interspecies transmission, including zoonotic transmission with bats as reservoir hosts. For the first time, a group 2 CoV was found in a bat, Pipistrellus pipistrellus, in Europe. This is of particular interest for public health as the reservoir host is a species that is common to urban areas in most of Europe and notorious for its close interactions with humans. Four verspertilionid bat species were found to excrete group 1 CoVs, viz. Myotis daubentonii, M. dasycneme, P. pipistrellus, and Nyctalus noctula. The last species is a newly identified reservoir. The overall prevalence was 16.9% and positive bats were found at multiple widespread locations. The circulating group 1 CoV lineages were rather species associated than location associated.


Cyclic oligothiophenes: novel organic materials and models for polythiophene. A theoretical study.
Cyclic oligothiophenes (CnT, n = 6-30, even only) in syn- and anti-conformations are studied theoretically at the B3LYP/6-31G(d) level of theory. Strain energies, ionization potentials, HOMO-LUMO gaps, bond length alternations, NICS values, and IR and Raman spectra have been studied. The properties of anti-conformers change systematically with increasing ring size and were studied in detail in neutral, radical cation, and dication forms. In syn-conformation, the oligomers lose their nearly planar ring shape and bend significantly for n > 14, and thus properties cannot be related to ring size. The HOMO-LUMO gap in C14T-syn is even lower than polythiopehene. IR and Raman spectra calculated at the B3LYP/6-31G(d) level are used to differentiate syn- from anti-conformations. The properties of cyclic oligomers are compared to those of the linear system, and cyclic oligothiophenes are revealed as good models for polythiophene. To assist the experimental study of known cyclic oligomers having dibutyl substituents on alternate thiophene rings, the corresponding dimethyl-substituted oligomers are also studied. The experimentally evaluated HOMO-LUMO gaps for alternately dibutyl-substituted cyclic oligomers match the calculated values; however, they are significantly higher than those of the unsubstituted analogues.


Fluorescent indolizine derivative YI-13 detects amyloid-β monomers, dimers, and plaques in the brain of 5XFAD Alzheimer transgenic mouse model.
Alzheimer disease (AD) is a neurodegenerative disorder characterized by the aberrant production and accumulation of amyloid-β (Aβ) peptides in the brain. Accumulated Aβ in soluble oligomer and insoluble plaque forms are considered to be a pathological culprit and biomarker of the disorder. Here, we report a fluorescent universal Aβ-indicator YI-13, 5-(4-fluorobenzoyl)-7,8-dihydropyrrolo[1,2-b]isoquinolin-9(6H)-one, which detects Aβ monomers, dimers, and plaques. We synthesized a library of 26 fluorescence chemicals with the indolizine core and screen them through a series of in vitro tests utilizing Aβ as a target and YI-13 was selected as the final imaging candidate. YI-13 was found to stain and visualize insoluble Aβ plaques in the brain tissue, of a transgenic mouse model with five familial AD mutations (5XFAD), by a histochemical approach and to label soluble Aβ oligomers within brain lysates of the mouse model under a fluorescence plate reader. Among oligomers aggregated from monomers and synthetic dimers from chemically conjugated monomers, YI-13 preferred the dimeric Aβ.


Characteristics of the thick, compound refractive lens.
A compound refractive lens (CRL), consisting of a series of N closely spaced lens elements each of which contributes a small fraction of the total focusing, can be used to focus x rays or neutrons. The thickness of a CRL can be comparable to its focal length, whereupon a thick-lens analysis must be performed. In contrast with the conventional optical lens, where the ray inside the lens follows a straight line, the ray inside the CRL is continually changing direction because of the multiple refracting surfaces. Thus the matrix representation for the thick CRL is quite different from that for the thick optical lens. Principal planes can be defined such that the thick-lens matrix can be converted to that of a thin lens. For a thick lens the focal length is greater than for a thin lens with the same lens curvature, but this lengthening effect is less for the CRL than for the conventional optical lens.


Could the results of posterior colporrhaphy and levatoroplasty be improved by abdominal sacrocolporectopexy?
The aim of our study was to improve poor results of preliminary colporrhaphy and anterior levatoroplasty performed for patients with low and middle rectocele. 55 patients who had undergone colporrhaphy and anterior levatoroplasty for rectocele in 2012- 2015 and contacted the clinic with complaints of ongoing obstructive defecation were included in this investigation. They had a comprehensive medical examination using defecography, ultrasound, magnetic resonance imaging, anorectal functional tests. This study revealed perineum descending in 20 patients, apical prolapse 23 patients, and 12 patients demonstrated simultaneous pathology. All these patients were performed additional abdominal sacrocolporectopexy in 2015-2017 and results were estimated. Abdominal sacrocolporectopexy significantly improved anatomical and functional results of previous surgery which was confirmed by the listed methods of research. The average location of the anorectal area and utero-cervical zone became higher. So, perineum descendents and apical prolapsed were improved. Sacrocolporectopexy is often used for surgical treatment of pelvic organ prolapse. Our study shows its efficiency in patients with perineum descending and upper rectocele. Abdominal sacrocolporectopexy is an effective method of surgical correction of relapses and unsatisfactory results of treatment of patients with pelvic prolapse, manifested by perineum body descending and apical prolapse. Apical prolapse, Levatoroplasty, Perineum descending, Rectocele, Sacrocolporectopexy.


Regulation of the gltBDF operon of Escherichia coli: how is a leucine-insensitive operon regulated by the leucine-responsive regulatory protein?
The regulon controlled by the leucine-responsive regulatory protein (Lrp) of Escherichia coli consists of over 40 genes and proteins whose expression is regulated, either positively or negatively, by Lrp. The gltBDF operon, encoding glutamate synthase, was originally identified as a member of the Lrp regulon through a two-dimensional electrophoretic analysis of polypeptides from isogenic strains containing or lacking a functional Lrp protein. We have now demonstrated that Lrp regulates the transcription of gltBDF::lacZ operon fusions. Relative to expression in glucose minimal 3-(N-morpholino)propanesulfonic acid (MOPS) medium, gltBDF::lacZ expression in an lrp+ strain is repressed 2.2-fold in the presence of 10 mM exogenous leucine and 16-fold in Luria broth. Repression of gltBDF::lacZ expression by leucine or Luria broth is not seen for an isogenic strain containing a Tn10 insertion in lrp, and expression of gltBDF::lacZ is 44-fold lower than in the lrp+ strain when both are grown in glucose minimal MOPS medium. Lrp binds specifically to DNA fragments containing the gltBDF promoter region. Saturating levels of leucine do not abolish binding of Lrp upstream of gltBDF but merely increase its apparent dissociation constant from 2.0 to 6.9 nM. Electrophoretic analysis of the Lrp regulon established that target proteins differ greatly in the degree to which the effect of Lrp on their expression is antagonized by leucine. On the basis of our present results, we present a model for positive regulation of target genes by Lrp. Insensitivity to leucine would be expected when the effective intracellular concentration of Lrp is high relative to the affinity of Lrp binding sites required for transcription of the target gene. At lower concentrations of Lrp, transcription of the target gene should be sensitive to leucine. This model suggests that regulation of the concentration of active Lrp is critical to control of the Lrp regulon.


The providers' coordination of care: a model for collaboration across the continuum of care.
With guidance from Health Services Advisory Group, the Arizona Medicare Quality Improvement Organization, a Prescott, Arizona, workgroup consisting of a hospital, nursing home, and 2 home health agencies have come together to improve continuity of care as related to pressure ulcer prevention and treatment. Weekly, then monthly, meetings were held to develop recommendations for practice standardization, a treatment grid for easy reference, an educational module with posttests, and standardized physicians' orders, which were developed and disseminated to the local provider community. More than 200 staff members were educated at Yavapai Regional Medical Center (YRMC), local skilled nursing facilities, and home health agencies. In addition, new employees and nursing students are now educated on the Providers' Coordination of Care Educational Module at YRMC. The Module has been placed on the hospital's Web site and offered free to all interested parties. With pressure being mounted by Centers for Medicare & Medicaid Services, the Office of the Inspector General (OIG), and the general public, the role of the case manager (in all practice settings) will increasingly become more crucial. Through development of a network of local venues of care, the case manager can facilitate care transitions throughout the healthcare system with the goal of improved outcomes for the patient regardless of setting. Providing the right care, in the right setting to improve patient care, can only benefit our aging and declining populations. Cross-setting work will become more essential as the Centers for Medicare & Medicaid Services goal of pay-for-performance is realized across all care settings. As a national priority, pressure ulcer prevention and treatment needs to be embraced by all settings. It serves the patient, as well as the healthcare providers, to improve care regardless of setting or competition. Positive results can be accomplished if the best interests of the patient are kept at the forefront. To this end, obstacles of competition and individual facility possessiveness must be overcome to achieve the desired results. Having the right mix of professionals will enhance the results. The lessons learned here can be used in any across-the-settings initiative.


Vaccination with a synthetic peptide modulates lymphocytic choriomeningitis virus-mediated immunopathology.
Vaccination with a nucleopeptide (NP 118; amino acids 118 to 132) representing a cytotoxic T-cell epitope of lymphocytic choriomeningitis virus (LCMV) can modulate immunopathology. Immunization with NP 118 protected H-2d mice against intracerebral infection with the LCMV-ARMSTRONG isolate. However, when NP 118-primed H-2d mice were challenged intracerebrally with an intermediate dose (5 x 10(4) PFU) of the LCMV-DOCILE strain, all mice primed with NP 118 emulsified in incomplete Freund's adjuvant died, whereas unprimed mice survived. Correspondingly, peptide vaccination enhanced specifically the cytotoxic T-cell response, influencing the critical balance between T-cell response and virus spread.


Experience with a low volume ultrafiltration cell in small children*.
A low volume (16 ml) ultrafiltration cell was used ten times in two small, fluid overloaded children to remove plasma water. The device was simple to use and, at slow blood flow rates (25-50 ml/minutes) and low transmembrane pressures (10-30 mm Hg), provided controlled removal of excess fluid. Although no major complications were encountered, hypothermia and hypotension (at ultrafiltrate flux rates exceeding 0.5 ml/kg/minute) were observed. The ultrafiltrate solute concentration was similar to plasma and no significant shifts in serum electrolytes were induced. The ultrafiltrate protein concentration of 64 to 2,760 mg/dl was much higher than previously reported.


NCBP2 modulates neurodevelopmental defects of the 3q29 deletion in Drosophila and Xenopus laevis models.
The 1.6 Mbp deletion on chromosome 3q29 is associated with a range of neurodevelopmental disorders, including schizophrenia, autism, microcephaly, and intellectual disability. Despite its importance towards neurodevelopment, the role of individual genes, genetic interactions, and disrupted biological mechanisms underlying the deletion have not been thoroughly characterized. Here, we used quantitative methods to assay Drosophila melanogaster and Xenopus laevis models with tissue-specific individual and pairwise knockdown of 14 homologs of genes within the 3q29 region. We identified developmental, cellular, and neuronal phenotypes for multiple homologs of 3q29 genes, potentially due to altered apoptosis and cell cycle mechanisms during development. Using the fly eye, we screened for 314 pairwise knockdowns of homologs of 3q29 genes and identified 44 interactions between pairs of homologs and 34 interactions with other neurodevelopmental genes. Interestingly, NCBP2 homologs in Drosophila (Cbp20) and X. laevis (ncbp2) enhanced the phenotypes of homologs of the other 3q29 genes, leading to significant increases in apoptosis that disrupted cellular organization and brain morphology. These cellular and neuronal defects were rescued with overexpression of the apoptosis inhibitors Diap1 and xiap in both models, suggesting that apoptosis is one of several potential biological mechanisms disrupted by the deletion. NCBP2 was also highly connected to other 3q29 genes in a human brain-specific interaction network, providing support for the relevance of our results towards the human deletion. Overall, our study suggests that NCBP2-mediated genetic interactions within the 3q29 region disrupt apoptosis and cell cycle mechanisms during development.


Multiple regulatory pathways associated with high-level ciprofloxacin and multidrug resistance in Salmonella enterica serovar enteritidis: involvement of RamA and other global regulators.
Mechanisms of antibiotic resistance were examined in nalidixic acid-resistant Salmonella enterica serovar Enteritidis field isolates displaying decreased susceptibility to ciprofloxacin and in in vitro-derived ciprofloxacin-resistant mutants (104-cip and 5408-cip). All field isolates harbored a single gyrA mutation (D87Y). Deletion of acrB and complementation with wild-type gyrA increased quinolone susceptibility. Selection for ciprofloxacin resistance was associated with the development of an additional gyrA (S83F) mutation in 104-cip, novel gyrB (E466D) and parE (V461G) mutations in 5408-cip, overexpression of acrB and decreased susceptibility to nonquinolone antibiotics in both mutants, and decreased OmpF production and altered lipopolysaccharide in 104-cip. Complementation of mutated gyrA and gyrB with wild-type alleles restored susceptibility to quinolones in 104-cip and significantly decreased the ciprofloxacin MIC in 5408-cip. Complementation of parE had no effect on quinolone MICs. Deletion of acrB restored susceptibility to ciprofloxacin and other antibiotics tested. Both soxS and marA were overexpressed in 104-cip, and ramA was overexpressed in 5408-cip. Inactivation of each of these global regulators lowered ciprofloxacin MICs, decreased expression of acrB, and restored susceptibility to other antibiotics. Mutations were found in soxR (R20H) and in soxS (E52K) in 104-cip and in ramR (G25A) in 5408-cip. In conclusion, both efflux activity and a single gyrA mutation contribute to nalidixic acid resistance and reduced ciprofloxacin sensitivity. Ciprofloxacin resistance and decreased susceptibility to multiple antibiotics can result from different genetic events leading to development of target gene mutations, increased efflux activity resulting from differential expression of global regulators associated with mutations in their regulatory genes, and possible altered membrane permeability.


Interferon-gamma induces biosynthesis of complement components C2, C4 and factor H by human proximal tubular epithelial cells.
In a previous study the authors demonstrated that the production of complement component C4 by human kidney proximal tubular epithelial cells (PTEC) is upregulated by interferon gamma (IFN-gamma). In the present study the authors describe that PTEC in culture express both mRNA and protein of the IFN-gamma receptor complex, and that culture of PTEC with 1000 U/ml IFN-gamma for 72 h results in enhanced production not only of C4 (36.1 ng/10(6) cells), but also of C2 (10.8 ng/10(6) cells) and Factor H (17.5 ng/10(6) cells). Unstimulated PTEC produced 0.5 ng/10(6) cells, 0.5 ng/10(6) cells and 0.4 ng/10(6) cells of C2, C4 and Factor H, respectively. The upregulation of the three complement components was dose- and time-dependent and specific for IFN-gamma because the effect of IFN-gamma was abolished by a monoclonal antibody directed against IFN-gamma. Furthermore no effect of other cytokines was observed. The regulation of synthesis of C2, C4 and Factor H occurred at the transcriptional level as shown by semi-quantitative RT-PCR and dot-blot analysis. Taken together with the observation that in normal kidney tissue the tubuli express IFN-gamma receptor alpha-chain and a signal transducing protein, the present study implies that enhanced production of complement by PTEC may occur during a local immune response by in situ generation of IFN-gamma by infiltrating T-cells in the interstitium of the kidney.


Rumination, Brooding, and Reflection: Prospective Associations with Suicide Ideation and Suicide Attempts.
Rumination is often cited as a risk factor for suicide, yet few studies of rumination have utilized clinical samples, and no studies have examined its prospective association with suicide attempts. The purpose of this study was to examine concurrent and prospective associations of brooding and reflection (the two components of rumination) with suicide ideation and suicide attempts among a high-risk clinical sample. Participants were 286 adolescents and young adults (77% Caucasian, 59% female) aged 13-25 seeking psychiatric emergency services. A majority (71%) were presenting with a primary complaint of suicide ideation or recent suicide attempt. Participants completed a baseline assessment at the index visit; 226 participants (79%) completed a 4-month follow-up assessment of suicidal thoughts and behaviors. Brooding was associated with lifetime history of one or more suicide attempts, but not concurrent suicide ideation. Reflection was not associated with lifetime suicide attempts or concurrent suicide ideation. Furthermore, prospective associations of brooding and reflection with suicide ideation and suicide attempts were weak-to-small in magnitude and statistically nonsignificant. Rumination appears to have a limited association with suicide-related outcomes within a high-risk clinical sample. Additional longitudinal studies utilizing clinical samples are critically needed to better understand these associations.


A specific RNA hairpin loop structure binds the RNA recognition motifs of the Drosophila SR protein B52.
B52, also known as SRp55, is a member of the Drosophila melanogaster SR protein family, a group of nuclear proteins that are both essential splicing factors and specific splicing regulators. Like most SR proteins, B52 contains two RNA recognition motifs in the N terminus and a C-terminal domain rich in serine-arginine dipeptide repeats. Since B52 is an essential protein and is expected to play a role in splicing a subset of Drosophila pre-mRNAs, its function is likely to be mediated by specific interactions with RNA. To investigate the RNA-binding specificity of B52, we isolated B52-binding RNAs by selection and amplification from a pool of random RNA sequences by using full-length B52 protein as the target. These RNAs contained a conserved consensus motif that constitutes the core of a secondary structural element predicted by energy minimization. Deletion and substitution mutations defined the B52-binding site on these RNAs as a hairpin loop structure covering about 20 nucleotides, which was confirmed by structure-specific enzymatic probing. Finally, we demonstrated that both RNA recognition motifs of B52 are required for RNA binding, while the RS domain is not involved in this interaction.


A liver-specific enhancer in the core promoter region of human hepatitis B virus.
An 88-base pair fragment in the core promoter of the human hepatitis B virus (HBV) contains a functional promoter and a strong liver-specific enhancer. This enhancer functions in human hepatoma cells, where it is much more active than the previously described HBV enhancer in stimulating expression of the linked bacterial chloramphenicol acetyltransferase gene expressed from heterologous promoters. Studies of the role of this enhancer-promoter in HBV may help to clarify mechanisms of gene expression in cells infected with HBV and the role of the virus in the pathogenesis of hepatitis and hepatocellular carcinoma.


Visual Performance of a New Extended Depth-of-Focus Intraocular Lens Compared to a Distance-Dominant Diffractive Multifocal Intraocular Lens.
To investigate the clinical performance of a new extended depth-of-focus (EDOF) intraocular lens (IOL) and compare it to that of a distance-dominant diffractive multifocal IOL. Patients implanted with an EDOF IOL (Mini Well; SIFI, Catania, Italy) inducing spherical aberration and with a multifocal IOL (ReSTOR SV25T; Alcon Laboratories, Inc., Fort Worth, TX) were analyzed. The following monocular parameters were investigated: corrected distance visual acuity (CDVA), distance-corrected near visual acuity (DCNVA), reading speed, defocus curve, contrast sensitivity, and halos and glare as quantified by a simulator (Halo & Glare Simulator; Eyeland-Design Network GmbH, Vreden, Germany) and questionnaire. Twenty patients with the EDOF IOL and 37 with the multifocal IOL were enrolled. No statistically significant difference was observed for CDVA. The defocus curve of the EDOF IOL revealed no gaps for the intermediate range. Statistically significant differences were observed at -1.00 diopter (D) (EDOF IOL: 0.08 ± 0.09 logMAR; multifocal IOL: 0.21 ± 0.12 logMAR; P < .0001) and -1.50 D defocus (EDOF IOL: 0.15 ± 0.11 logMAR; multifocal IOL: 0.24 ± 0.13 logMAR; P = .0122). The reading speed at 40 cm was similar at all print sizes. The mean DCNVA was the same (EDOF IOL: 0.35 ± 0.14 logRAD, multifocal IOL: 0.35 ± 0.13 logRAD). No differences in contrast sensitivity were detected. According to the simulator, halos had a smaller mean size (P = .0439) and a lower mean intensity (P = .0222) with the EDOF IOL. No statistically significant differences were detected for glare size. The new EDOF IOL performed similarly to a multifocal IOL at distance and near but was superior at intermediate distances. [J Refract Surg. 2018;34(4):228-235.].


Continuous variation rather than specialization in the egg phenotypes of cuckoos (Cuculus canorus) parasitizing two sympatric reed warbler species.
The evolution of brood parasitism has long attracted considerable attention among behavioural ecologists, especially in the common cuckoo system. Common cuckoos (Cuculus canorus) are obligatory brood parasites, laying eggs in nests of passerines and specializing on specific host species. Specialized races of cuckoos are genetically distinct. Often in a given area, cuckoos encounter multiple hosts showing substantial variation in egg morphology. Exploiting different hosts should lead to egg-phenotype specialization in cuckoos to match egg phenotypes of the hosts. Here we test this assumption using a wild population of two sympatrically occurring host species: the great reed warbler (Acrocephalus arundinaceus) and reed warbler (A. scirpaceus). Using colour spectrophotometry, egg shell dynamometry and egg size measurements, we studied egg morphologies of cuckoos parasitizing these two hosts. In spite of observing clear differences between host egg phenotypes, we found no clear differences in cuckoo egg morphologies. Interestingly, although chromatically cuckoo eggs were more similar to reed warbler eggs, after taking into account achromatic differences, cuckoo eggs seemed to be equally similar to both host species. We hypothesize that such pattern may represent an initial stage of an averaging strategy of cuckoos, that--instead of specializing for specific hosts or exploiting only one host--adapt to multiple hosts.


[Lesions of the renal pedicle in blunt abdominal trauma in children. 7 cases].
The frequency of urinary tract trauma in children is difficult to estimate. Renal pedicle lesions are estimated to represent 10% of all forms of urinary tract trauma. These lesions are difficult to diagnose and may be completely missed. Over a 12 year period, we have treated 7 children with a renal pedicle injury. Haematuria was immediate in 3 cases, delayed in 2 cases and absent in 2 cases. The diagnosis is based on intravenous urography or postcontrast computed tomography. Treatment consisted of disobstruction with intimal suture with a good result, but impaired renal function, an upper pole nephrectomy, functional aorto-renal bypass graft but with altered renal function, total nephrectomy and 3 cases of surveillance with one non-functioning kidney. None of these children are hypertensive. The authors discuss the diagnostic approach, the contribution of imaging and the therapeutic options and results.


PEGylated and Non-PEGylated TCP-1 Probes for Imaging of Colorectal Cancer.
Previous studies indicate that 99mTc- and fluorescent-labeled c[Cys-Thr-Pro-Ser-Pro-Phe-Ser-His-Cys]OH (TCP-1) peptides were able to detect colorectal cancer (CRC) and tumor-associated vasculature. This study was designed to characterize the targeting properties of PEGylated and non-PEGylated TCP-1 peptides for CRC imaging. Cell uptake of cyanine 7 (Cy7)-labeled TCP-1 probes (Cy7-PEG4-TCP-1 and Cy7-TCP-1) was investigated in three CRC cell lines (human, HCT116 and HT29; mouse, CT26). Xenograft and orthotopic CRC tumor models with HCT116 and CT26 cells were used to characterize biodistribution and CRC tumor-targeting properties of TCP-1 fluorescence and radioligand with and without PEGylation, [99mTc]Tc-HYNIC-PEG4-TCP-1 vs. [99mTc]Tc-HYNIC-TCP-1. Fluorescence images showed that TCP-1 probes were distributed in the cytoplasm and nucleus of CRC cells. When CT26 cells were treated with unlabeled TCP-1 peptide prior to the cell incubation with Cy7-PEG4-TCP-1, cell fluorescent signals were significantly reduced relative to the cells without blockade. Relative to Cy7-TCP-1, superior brilliance and visibility of fluorescence was observed in the tumor with Cy7-PEG4-TCP-1 and maintained up to 18 h post-injection. [99mTc]Tc-HYNIC-PEG4-TCP-1 images in xenograft and orthotopic CRC models demonstrated that TCP-1 PEGylation preserved tumor-targeting capability of TCP-1, but its distribution (%ID/g) in the liver and intestine was higher than that of [99mTc]Tc-HYNIC-TCP-1 (1.51 ± 0.29 vs 0.53 ± 0.12, P < 0.01). Better tumor visualization by [99mTc]Tc-HYNIC-TCP-1 was observed in the orthotopic CRC model due to lower intestinal radioactivity. TCP-1-based probes undergo endocytosis and localize in the cytoplasm and nucleus of human and mouse CRC cells. Tumor detectability of fluorescent TCP-1 peptide with a PEG4 spacer is promising due to its enhanced tumor binding affinity and rapid clearance kinetics from nontumor tissues. Non-PEGylated [99mTc]Tc-HYNIC-TCP-1 exhibits lower nonspecific accumulation in the liver and gastrointestinal tract and might have better capability for detecting CRC lesions in clinical sites. TCP-1 may represent an innovative targeting molecule for detecting CRC noninvasively.


Fibroblast PER2 circadian rhythmicity depends on cell density.
Like neurons in the suprachiasmatic nucleus (SCN), the master circadian pacemaker in the brain, single fibroblasts can function as independent oscillators. In the SCN, synaptic and paracrine signaling among cells creates a robust, synchronized circadian oscillation, whereas there is no evidence for such integration in fibroblast cultures. However, interactions among single-cell fibroblast oscillators cannot be completely excluded, because fibroblasts were not isolated in previous work. In this study, we tested the autonomy of fibroblasts as single-cell circadian oscillators in high- and low-density culture, by single-cell imaging of cells from PER2::LUC circadian reporter mice. We found greatly reduced PER2::LUC rhythmicity in low-density cultures, which could result from lack of either constitutive or rhythmic paracrine signals from neighboring fibroblasts. To discriminate between these 2 possibilities, we mixed PER2::LUC wild-type (WT) cells with nonluminescent, nonrhythmic Bmal1-/- cells, so that density of rhythmic cells was low but overall cell density remained high. In this condition, WT cells showed clear rhythmicity similar to high-density cultures. We also mixed PER2::LUC WT cells with nonluminescent, long period Cry2-/- cells. In this condition, WT cells showed a period no different from cells cultured with rhythmic WT cells or nonrhythmic Bmal1-/- cells. In previous work, we found that low K⁺ suppresses fibroblast rhythmicity, and we and others have found that either low K⁺ or low Ca²⁺ suppresses SCN rhythmicity. Therefore, we attempted to rescue rhythmicity of low-density fibroblasts with high K⁺ (21 mM), high Ca²⁺ (3.6 mM), or conditioned medium. Conditioned medium from high-density fibroblast cultures rescued rhythmicity of low-density cultures, whereas high K⁺ or Ca²⁺ medium did not consistently rescue rhythmicity. These data suggest that fibroblasts require paracrine signals from adjacent cells for normal expression of rhythmicity, but that these signals do not have to be rhythmic, and that rhythmic signals from other cells do not affect the intrinsic periods of fibroblasts.


Generic hyper-diversity in Stachybotriaceae.
The family Stachybotriaceae was recently introduced to include the genera Myrothecium, Peethambara and Stachybotrys. Members of this family include important plant and human pathogens, as well as several species used in industrial and commercial applications as biodegraders and biocontrol agents. However, the generic boundaries in Stachybotriaceae are still poorly defined, as type material and sequence data are not readily available for taxonomic studies. To address this issue, we performed multi-locus phylogenetic analyses using partial gene sequences of the 28S large subunit (LSU), the internal transcribed spacer regions and intervening 5.8S nrRNA (ITS), the RNA polymerase II second largest subunit (rpb2), calmodulin (cmdA), translation elongation factor 1-alpha (tef1) and β-tubulin (tub2) for all available type and authentic strains. Supported by morphological characters these data resolved 33 genera in the Stachybotriaceae. These included the nine already established genera Albosynnema, Alfaria, Didymostilbe, Myrothecium, Parasarcopodium, Peethambara, Septomyrothecium, Stachybotrys and Xepicula. At the same time the generic names Melanopsamma, Memnoniella and Virgatospora were resurrected. Phylogenetic inference further showed that both the genera Myrothecium and Stachybotrys are polyphyletic resulting in the introduction of 13 new genera with myrothecium-like morphology and eight new genera with stachybotrys-like morphology.


From the transnational to the Sinophone: lesbian representations in Chinese-language films.
This article theorizes global lesbian cinema in Chinese-language films through regionalism, diaspora studies, and Sinophone studies. Through an inter-regional analysis of Butterfly (Yan Yan Mak, 2004, Hong Kong) and diasporic and Sinophone readings of Saving Face (Alice Wu, 2005, USA), I argue that Mak's film illustrates a Hong Kong regional retranslation of a Taiwanese lesbian story, which complicates any claim to a stable "Chinese" identity. Finally, Wu's representation of lesbianism also troubles the politics of Chineseness by pointing to the ways diasporic reproduction of "community" works through the disciplining of other non-normative sexualities.


Stridulation in Leaf-Cutting Ants.
The leaf-cutting ant Atta caphaloes L. stridulates whenever it is prevented from moving freely. Although audible to the human ear, the airborne sound produced has its main energy concentrated between 20 and 60 kilocycles per second. However, it is not the airborne, but the groundconducted stridulation sound that acts as a distress alarm: a stridulating ant attracts other workers, and if the "calling" ant is covered by earth, intensive digging is released in the attracted nest mates.


Thoracoscopic esophagectomy: are there benefits?
Between 1991 and 1995, 18 patients affected by a resectable intramural tumor of the esophagus underwent esophagectomy with thoracoscopic dissection of the esophagus. All patients had a relative contraindication to transthoracic esophagectomy with radical lymphadenectomy. All esophagectomies were completed thoracoscopically and reconstruction of the digestive tract was performed in 17 cases through cervical gastroplasty, and in 1 case, through cervical coloplasty. One cirrhotic patient died in the postoperative period due to a cervical anastomotic leak. Six other patients experienced a postoperative complication (mortality rate, 5.5%; morbidity rate, 33.3%). After a median follow-up of 17 months, 14 patients are alive without evidence of disease. One patient, who had excision of a cutaneous metastasis at a trocar insertion site 6 months postoperatively, eventually died with locoregional recurrence 14 months postoperatively. Another patient died 20 months after surgery with mediastinal recurrence. One patient died 28 months postoperatively after massive hematemesis with a suspect abdominal recurrence. The results of the present series, and those reported by other authors, do not seem to indicate evident advantages at present for the minimally invasive procedure during resection of the esophagus for cancer. Currently, there is no indication that this procedure should be used for standard clinical use. Wider randomized trials, performed in selected centers only, and longer follow-up are needed to further evaluate the procedure.


Distinct histopathological patterns in single lesion leprosy patients treated with single dose therapy (ROM) in the Brazilian Multicentric Study.
This paper aims to describe the histomorphologic features of skin biopsies of single lesion leprosy patients recruited at outpatient clinics in four Brazilian states in the Northeast (Amazonas and Rondonia), Southeast (Rio de Janeiro) and Center-West (Goiás) between October 1997 and December 1998. Patients clinically diagnosed as single skin lesion paucibacillary (SSL-PB) leprosy had a standard 4-mm punch biopsy taken from the lesion before rifampin, ofloxacin, minocycline (ROM) therapy. The features of the cellular inflammatory infiltrates, the presence of nerve involvement and acid-fast bacilli (AFB) were used to categorize SSL-PB biopsies into different histopathological groups. Two-hundred-seventy-eight (93.0%) out of 299 patients had a skin biopsy available. Seven single lesion patients were diagnosed as BL or LL leprosy types (MB) by the histopathological exams and 12 cases were excluded due to other skin diseases. Therefore, 259 patients had skin lesions with histomorphological features compatible with PB leprosy categorized as follows: 33.6% (N = 87) of the biopsies represented well-circumscribed epithelioid cell granuloma (Group 1); 21.6% (N = 56) less-circumscribed epithelioid cell granuloma (Group 2); 12.0% (N = 31) were described as mononuclear inflammatory infiltrate permeated with epithelioid cells (Group 3), and 29.7% (N = 77) had perivascular/periadnexal mononuclear inflammatory infiltrate (Group 4). Minimal/no morphological alteration in the skin was detected in only 8 (3.1%) SSL-PB patients categorized as Group 5, who were considered to have leprosy by clinical parameters. SSL-PB leprosy patients recruited in a multicentric study presented histomorphology readings comprising the whole PB leprosy spectrum but also a few MB cases. These results indicate heterogeneity among SSL-PB patients, with a predominance of well-circumscribed and less-circumscribed epithelioid cell granulomas (Groups 1 and 2) in the sites studied and the heterogeneity of local cellular immune response.


Laparoscopic management of peritoneal dialysis catheters.
Laparoscopic surgical techniques were used for peritoneal dialysis catheter placement or for revision in patients with previous extensive abdominal surgical treatment, multiple prior catheter insertions or dysfunctional catheters with outflow obstruction. Nineteen laparoscopic procedures were performed upon 16 patients between July 1990 and January 1992. All patients were chosen for the laparoscopic approach because of the complicated nature of previous incisions or in an attempt to salvage existing catheters. In five patients, open laparoscopy was performed with the blunt Hasson trocar, and in the remaining patients, closed techniques were used to establish the pneumoperitoneum. Dysfunctional catheters were revised by adhesiolysis, partial omental resection and subtotal omentectomy. New catheters were easily placed percutaneously, sometimes after extensive adhesiolysis, using direct laparoscopic guidance. All patients had functioning catheters postoperatively. Two catheters subsequently failed because of dislodgement and recurrent obstruction. The overall success rate for the entire series was 75 percent. We conclude that laparoscopy will be a useful addition to the surgical armamentarium in managing patients on peritoneal dialysis, particularly as this mode of renal replacement therapy rapidly expands.


Familiar communication partners' facilitation of topic management in conversations with individuals with dementia.
Language and memory impairments affect everyday interactions between individuals with dementia and their communication partners. Impaired topic management, which compromises individuals' construction of relevant, meaningful discourse, is commonly reported amongst individuals with dementia. Currently, limited empirical evidence describes the sequential patterns of behaviour comprising topic-management practices in everyday conversation between individuals with dementia and their communication partners. To describe the sequential patterns of behaviour relating to the manifestation of topic-management impairments and facilitative behaviours in everyday interactions between individuals with dementia and their familiar communication partners (FCPs). Three 20-min conversations between individuals with moderate to severe dementia and their FCPs were recorded. Conversation Analysis was used to examine sequences in which topic-management appeared to be impaired. Conversational behaviours that reflected a difficulty in contributing on-topic talk were pervasive in the talk of the three individuals with dementia. FCPs responded to these conversational difficulties by using two categories of facilitative behaviours. The first involved responding to an individual with dementia's explicit repair-initiation by performing repair. In the second category, explicit repair-initiation was absent; instead, the distance of the conversational difficulty from the prior topic-shifting turn mediated the form and outcome of the FCPs' facilitative behaviours. Each category successfully facilitated the individual with dementia to contribute on-topic talk. The findings contribute to a growing understanding of topic-management abilities in everyday interactions involving individuals with dementia. Individuals with dementia took a proactive role in eliciting topic-management support. The FCPs responded with turns that facilitated the individuals with dementia to talk on-topic. Clinically, the results support and extend the current topic-management recommendations available in communication partner training programmes, and promote conversations which attend to the personhood of the individual with dementia.


Disruption of gastrulation movements in Xenopus by a dominant-negative mutant for C-cadherin.
Gastrulation in Xenopus laevis transforms a ball of undifferentiated cells into a well-organized elongated embryo with a distinct body axis, as a result of extensive cell rearrangements and movements. Since cell adhesion is thought to play an important role during tissue morphogenesis, we investigated the potential role for C-cadherin, a primary cell-cell adhesion molecule in Xenopus, in gastrulation movements. A deletion mutant consisting of the extracellular and transmembrane domains but lacking the cytoplasmic tail (Ctrunc) was constructed to act as a dominant-negative inhibitor of the endogenous protein. Injection of in vitro transcribed Ctrunc mRNA into the prospective dorsal involuting marginal zone, the region that undergoes the most extensive movements, caused a reproducible defect in gastrulation. The defect, an inability to complete involution and close the blastopore, was cadherin-specific, because it could be rescued by expressing full length C-cadherin. Ctrune also seems to inhibit morphogenetic movements of animal cap explants isolated from injected embryos. Expression of Ctrunc did not appear to interfere with inductive processes in the embryos that developed gastrulation defects, because they formed complete axial structures, including a complete head and notochord. These results demonstrate that C-cadherin plays a critical role in Xenopus laevis gastrulation movements, and they support the hypothesis that cadherins can mediate cellular rearrangements during tissue morphogenesis.


Local recurrence and survival following nerve-sparing radical cystoprostatectomy.
From March 1982 through July 1988, 76 men underwent nerve-sparing radical cystoprostatectomy for carcinoma of the bladder at our hospital. Of the 76 patients 2 (2.6%) had positive surgical margins (dome of the bladder and left ureter) and neither had positive margins at the site of nerve-sparing modifications. Of 3 patients (3.9%) who had local recurrence none had positive surgical margins. The 5-year actuarial local recurrence rate is 7.5%. Thirteen of 76 patients (17%) died of transitional cell carcinoma and 7 (9%) died of other causes, while 53 (70%) are alive without evidence of disease with a mean followup of 38.4 months. The 5-year actuarial survival rates are 64% over-all, 68% without disease and 78% disease-specific. Of the 42 evaluable men who underwent cystoprostatectomy alone 27 (64%) are potent, compared to 2 of the 12 men (17%) who also underwent urethrectomy. We conclude that the nerve-sparing modifications do not compromise cancer control, that local recurrence and survival rates are at least comparable to those achieved with standard radical cystoprostatectomy, and that it is possible to preserve potency in most men undergoing this procedure.


[Personal experience in surgical resection of pancreatic carcinoma].
The period 1981-1993, 124 patients underwent resection for pancreatic cancer; they represented 30.8% of all patients observed in that period. Surgical procedures were a pancreatoduodenectomy (PD) in 100 cases, a total pancreatectomy (TP) in 3 cases and a distal pancreatectomy in 21 cases. PD was performed by the injection of Neoprene into the residual pancreatic stump, rather than of pancreatojejunal anastomosis; in 37 cases a pylorus-preserving PD was performed. Adjuvant treatments were given in addition to resection in 75 patients: 47 underwent intraoperative radiation therapy, following a PD in 41 cases, a TP in 1 case and a distal pancreatectomy in 5 cases. Overall operative mortality was 2.4%, overall morbidity 26.6%. In the 103 patients undergoing PD or TP mortality was 2.9% and morbidity 29.1%. Postoperative diabetes occurred in 12.8% of patients undergoing PD with Neoprene injection. Overall median survival was 16 months. Survival was significantly related to UICC staging distribution, to the radical nature of the operation or to adjuvant treatments. No relationship was found between survival and the type of resection (PD versus distal pancreatectomy) or the pylorus preservation. Pancreatic resection, whenever technically possible, represents the treatment of choice of localised pancreatic cancer, in association with adjuvant treatments.


Identification of potentially cytotoxic lesions induced by UVA photoactivation of DNA 4-thiothymidine in human cells.
Photochemotherapy-in which a photosensitizing drug is combined with ultraviolet or visible radiation-has proven therapeutic effectiveness. Existing approaches have drawbacks, however, and there is a clinical need to develop alternatives offering improved target cell selectivity. DNA substitution by 4-thiothymidine (S(4)TdR) sensitizes cells to killing by ultraviolet A (UVA) radiation. Here, we demonstrate that UVA photoactivation of DNA S(4)TdR does not generate reactive oxygen or cause direct DNA breakage and is only minimally mutagenic. In an organotypic human skin model, UVA penetration is sufficiently robust to kill S(4)TdR-photosensitized epidermal cells. We have investigated the DNA lesions responsible for toxicity. Although thymidine is the predominant UVA photoproduct of S(4)TdR in dilute solution, more complex lesions are formed when S(4)TdR-containing oligonucleotides are irradiated. One of these, a thietane/S(5)-(6-4)T:T, is structurally related to the (6-4) pyrimidine:pyrimidone [(6-4) Py:Py] photoproducts induced by UVB/C radiation. These lesions are detectable in DNA from S(4)TdR/UVA-treated cells and are excised from DNA more efficiently by keratinocytes than by leukaemia cells. UVA irradiation also induces DNA interstrand crosslinking of S(4)TdR-containing duplex oligonucleotides. Cells defective in repairing (6-4) Py:Py DNA adducts or processing DNA crosslinks are extremely sensitive to S(4)TdR/UVA indicating that these lesions contribute significantly to S(4)TdR/UVA cytotoxicity.


Rat pancreatic lipase and two related proteins: enzymatic properties and mRNA expression during development.
We report the cDNA sequences of rat colipase, rat pancreatic lipase (rPL), and a rat pancreatic lipase-related protein (rPLRP). Comparison to the human PLRP cDNA suggests that the isolated clone encodes rPLRP-2. Both cDNA and a third cDNA encoding rPLRP-1 are secreted from Sf9 cells infected with recombinant baculovirus. rPL and rPLRP-2 hydrolyze triolein, 8.0 and 4.4 mumol.min-1.microgram-1, respectively. They are inhibited by bile salts, and activity is restored by (pro)colipase. PLRP-1 has barely detectable activity against triolein, even with (pro)colipase present. The pattern of mRNA expression during development in the rat reveals that all mRNA are low in the fetal rat pancreas. Both PLRP mRNA rise just before birth to a maximum 12 h after birth. They fall to low levels in the adult. In contrast, the PL mRNA is low at birth and rises rapidly during the suckling-weanling transition. In conclusion, the rat has at least three genes encoding different lipases, and these related genes have separate regulatory controls.


Lysophosphatidic acid triggers inflammation in the liver and white adipose tissue in rat models of 1-acyl-sn-glycerol-3-phosphate acyltransferase 2 deficiency and overnutrition.
AGPAT2 (1-acyl-sn-glycerol-3-phosphate-acyltransferase-2) converts lysophosphatidic acid (LPA) into phosphatidic acid (PA), and mutations of the AGPAT2 gene cause the most common form of congenital generalized lipodystrophy which leads to steatohepatitis. The underlying mechanism by which AGPAT2 deficiency leads to lipodystrophy and steatohepatitis has not been elucidated. We addressed this question using an antisense oligonucleotide (ASO) to knockdown expression of Agpat2 in the liver and white adipose tissue (WAT) of adult male Sprague-Dawley rats. Agpat2 ASO treatment induced lipodystrophy and inflammation in WAT and the liver, which was associated with increased LPA content in both tissues, whereas PA content was unchanged. We found that a controlled-release mitochondrial protonophore (CRMP) prevented LPA accumulation and inflammation in WAT whereas an ASO against glycerol-3-phosphate acyltransferase, mitochondrial (Gpam) prevented LPA content and inflammation in the liver in Agpat2 ASO-treated rats. In addition, we show that overnutrition, due to high sucrose feeding, resulted in increased hepatic LPA content and increased activated macrophage content which were both abrogated with Gpam ASO treatment. Taken together, these data identify LPA as a key mediator of liver and WAT inflammation and lipodystrophy due to AGPAT2 deficiency as well as liver inflammation due to overnutrition and identify LPA as a potential therapeutic target to ameliorate these conditions.


Intraseptal microinjections of substance P and analogs potentiate pentobarbital-induced narcosis and depression of hippocampal cholinergic activity.
Intraseptal microinjection of Substance P (SP) has been shown to depress activity in the septal-hippocampal cholinergic pathway in the rat. Pentobarbital also depresses septal-hippocampal cholinergic activity, and a relationship between this depressed activity and pentobarbital-induced narcosis is suggested by a variety of studies. To examine this relationship further, we microinjected SP and its analogs, [D-Arg1, D-Pro2, D-Trp7,9, Leu11]-SP and [D-Pro2, D-Trp7,9]-SP, intraseptally in rats pretreated with pentobarbital, and measured the duration of loss of righting reflex and change in choline uptake in hippocampal synaptosomes. The duration of pentobarbital-induced loss of righting reflex was prolonged and the pentobarbital-induced reduction of hippocampal choline uptake was enhanced by all three drugs. A negative correlation (r = -0.96, P less than .02) was seen between duration of loss of righting reflex and synaptosomal choline uptake. Thus, the two analogs of SP appear to act as SP agonists at the septum, judged by their abilities to potentiate pentobarbital narcosis and reduce septal-hippocampal cholinergic activity. This is in contrast to reported actions of these two analogs as SP antagonists in various peripheral tissues.


[HER2 testing for breast cancer. Swedish laboratories can now offer a quality assured analysis of growth factor].
Between 20 and 30 per cent of all breast cancers show an overexpression of the growth factor HER2 and this implies a worse prognosis. Metastasising breast cancer that overexpress HER2 can effectively be treated with trastuzumab (Herceptin). In order to determine whether this treatment is indicated, HER2 expression must be quantified with immunohistochemical or molecular biological methods. Several factors may influence the laboratory analysis of HER2, such as choice of antibodies and probes, the reliability of the test performance itself and the performance of the pathologist(s) interpreting the test. National guidelines for HER2 testing have been published in several countries. In this article a series of quality assurance projects leading to the formulation of Swedish guidelines for HER2 testing is described. These projects have demonstrated that several Swedish pathology laboratories can reliably perform and interpret analysis of HER2 status today.


Development of a genomics module within an epilepsy-specific electronic health record: Toward genomic medicine in epilepsy care.
Both clinical genomics and e-Health technology are changing the way medicine is being practiced. Although the basic clinical methodology of good medical care will remain unchanged, the combined power of genomics and electronic health records has the capability of enhancing, and in some cases transforming, the practice of medicine. This is particularly true in the care of patients with complex long-term medical conditions such as chronic refractory epilepsy, especially in those with related complex comorbidities including intellectual disability and psychiatric disease. Herein we outline the development and integration of an epilepsy genomics module into a preexisting epilepsy electronic patient record (EPR) system. We describe how this EPR infrastructure is used to facilitate discussion at multidisciplinary clinical meetings around molecular diagnosis and resulting changes in management. This work illustrates the role of eHealth technology in embedding genomics into the clinical pathway.


NAD metabolism modulates inflammation and mitochondria function in diabetic kidney disease.
Diabetes mellitus is the leading cause of cardiovascular and renal disease in the United -States. Despite the beneficial interventions available for patients with diabetes, there remains a need for additional therapeutic targets and therapies in diabetic kidney disease (DKD). Inflammation and oxidative stress are increasingly recognized as important causes of renal diseases. Inflammation is closely associated with mitochondrial damage. The molecular connection between inflammation and mitochondrial metabolism remains to be elucidated. Recently, nicotinamide adenine nucleotide (NAD+) metabolism has been found to regulate immune function and inflammation. In the present studies, we tested the hypothesis that enhancing NAD metabolism could prevent inflammation in and progression of DKD. We found that treatment of db/db mice with type 2 diabetes with nicotinamide riboside (NR) prevented several manifestations of kidney dysfunction (i.e., albuminuria, increased urinary kidney injury marker-1 (KIM1) excretion, and pathologic changes). These effects were associated with decreased inflammation, at least in part via inhibiting the activation of the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) signaling pathway. An antagonist of the serum stimulator of interferon genes (STING) and whole-body STING deletion in diabetic mice showed similar renoprotection. Further analysis found that NR increased SIRT3 activity and improved mitochondrial function, which led to decreased mitochondrial DNA damage, a trigger for mitochondrial DNA leakage which activates the cGAS-STING pathway. Overall, these data show that NR supplementation boosted NAD metabolism to augment mitochondrial function, reducing inflammation and thereby preventing the progression of diabetic kidney disease.