A microscope stage sample holder for microfluorometric measurements of biological specimen in the range of temperature between 3.5 to 300 K.
A cryostat was constructed, which permits measurements of the intracellular fluorescence within a range of temperature between 3.5 to 300 K. The cooling unit operates in accordance with the "Continuous Flow Principle" and allows the application of objectives up to a numerical aperture of 0.6. It results from measurement of BAO Feulgen stained nuclear DNA, that a decrease of fluorescence intensity is caused by two different mechanisms: (1) There is a highly temperature dependent effect, originating from phenomena of solid physics, and (2) a second effect, which is almost temperature independent, and can be explained as a photochemical reaction.


Selective effector coupling of muscarinic acetylcholine receptor subtypes.
Attempts have been made by means of recombinant DNA technology to understand the molecular basis of the functional heterogeneity of the muscarinic acetylcholine receptor (mAChR). Molecularly defined mAChR subtypes have been produced from the cloned DNAs in Xenopus oocytes and NG108-15 neuroblastoma-glioma hybrid cells as transient and stable expression systems, respectively, and agonist-induced cellular responses have been examined. The results obtained provide evidence that mAChR subtypes are selectively coupled with different effector systems, albeit not exclusively.


Fast and simple methods for the optimization of kurtosis used as a projection pursuit index.
As a powerful method for exploratory data analysis, projection pursuit (PP) often outperforms principal component analysis (PCA) to discover important data structure. PP was proposed in 1970s but has not been widely used in chemistry largely because of the difficulty in the optimization of projection indices. In this work, new algorithms, referred as "quasi-power methods", are proposed to optimize kurtosis as a projection index. The new algorithms are simple, fast, and stable, which makes the search for the global optimum more efficient in the presence of multiple local optima. Maximization of kurtosis is helpful in the detection of outliers, while minimization tends to reveal clusters in the data, so the ability to search separately for the maximum and minimum of kurtosis is desirable. The proposed algorithms can search for either with only minor changes. Unlike other methods, no optimization of step size is required and sphering or whitening of the data is not necessary. Both univariate and multivariate kurtosis can be optimized by the proposed algorithms. The performance of the algorithms is evaluated using three simulated data sets and its utility is demonstrated with three experimental data sets relevant to analytical chemistry.


Appendectomy and risk of Parkinson's disease in two large prospective cohorts of men and women.
Prior work on appendectomy and PD has produced mixed results. In this study we examined whether history of self-reported appendectomy was related to risk of incident Parkinson's disease in the Nurses' Health Study and the Health Professionals Follow-up Study. We used the Cox proportional hazards model to estimate the hazard ratio of Parkinson's disease associated with self-report of appendectomy in men and women. Among women, we estimated the hazard ratio of Parkinson's disease associated with appendectomy for appendicitis and incidental appendectomy. In pooled analyses, self-report of any appendectomy was not related to Parkinson's disease risk: the hazard ratio of Parkinson's disease comparing participants who reported any appendectomy with those who did not was 1.08 (95% confidence interval, 0.94-1.23). In women, appendectomy for appendicitis, but not incidental appendectomy, was associated with a modestly elevated risk of Parkinson's disease (hazard ratio, 1.23 [95% confidence interval, 1.00-1.50]). Overall, this study suggests limited to no association between appendectomy and Parkinson's disease risk. © 2018 International Parkinson and Movement Disorder Society.


Gut-origin sepsis: evolution of a concept.
The concept of bacterial translocation and gut-origin sepsis as a cause of systemic infectious complications and the multiple organ dysfunction syndrome (MODS) in surgical and ICU patients has emerged over the last several decades, although the exact clinical relevance of these phenomena continues to be debated. Thus, the goal of this review is to trace the evolution of gut-origin sepsis and gut-induced MODS and put these disorders and observations into clinical perspective. Additionally, the mechanisms leading to gut-derived complications are explored as well as therapeutic options to limit or prevent these complications. From this work, several major conclusions emerge. First, that bacterial translocation occurs clinically and is responsible for increased infectious complications in patients undergoing major abdominal surgery. However, the phenomenon of bacterial translocation is not sufficient to explain the development of MODS in ICU patients. Instead, the development of MODS in these high-risk patients is likely due to gut injury and the systemic spread of non-microbial, tissue-injurious factors that reach the systemic circulation via the intestinal lymphatics. These observations have resulted in the gut-lymph hypothesis of MODS.


Direct evidence for the contribution of the unique I-ANOD to the development of insulitis in non-obese diabetic mice.
Insulin-dependent diabetes mellitus is characterized by the infiltration of lymphocytes into the islets of Langerhans of the pancreas (insulitis) followed by destruction of insulin-secreting beta-cells leading to overt diabetes. The best model for the disease is the non-obese diabetic (NOD) mouse. Two unusual features of the class II major histocompatibility complex (MHC) of the NOD mouse are the absence of I-E and the presence of unique I-A molecules (I-ANOD), in which aspartic acid at position 57 of the beta-chain is replaced by serine. This feature is also found in the HLA-DQ chain of many Caucasians with insulin-dependent diabetes mellitus. We have previously reported that the expression of I-E prevents the development of insulitis in NOD mouse. Here we report that the expression of I-Ak (A alpha kA beta k) in transgenic NOD mice can also prevent insulitis, and that this protection is seen not only when the I-A beta-chain has aspartic acid as residue 57, but also when this residue is serine. These results show that the single amino-acid substitution at position 57 of the I-A beta-chain from aspartic acid to serine is not sufficient for the development of the disease.


Perioperative administration of dexamethasone to prevent postoperative shivering: a systematic review and meta-analysis of randomized controlled trials.
To evaluate the perioperative administration of dexamethasone to prevent postoperative shivering. We searched PubMed, Embase, Google Scholar, Web of Science, and Cochrane Library for relevant studies of the administration of dexamethasone to prevent postoperative shivering published through 31 May 2023. The primary outcome was the incidence of postoperative shivering. Secondary outcomes comprised the incidence of postoperative nausea, vomiting, and postoperative nausea and vomiting (PONV). RevMan 5.3 software was used for the data analysis. We included 12 randomized controlled trials (1276 participants). The results revealed a benefit favoring the perioperative administration of dexamethasone to prevent postoperative shivering (relative risk [RR]: 0.39; 95% confidence interval [CI]: 0.23-0.63), as well as the grade of shivering. The administration of dexamethasone also reduced the incidence of postoperative nausea (RR: 0.54; 95% CI: 0.39-0.73), postoperative vomiting (RR: 0.37; 95% CI: 0.20-0.65), and PONV (RR: 0.50; 95% CI: 0.26-0.95) compared with the control group. This study indicated that perioperative administration of dexamethasone prevented postoperative shivering and decreased the incidence of other complications.PROSPERO registration number: CRD42020164488.


Gender-Based Discrimination Among Medical Students: A Cross-Sectional Study in Brazil.
Gender-based discrimination (GBD) creates a hostile environment during medical school, affecting students' personal life and academic performance. Little is known about how GBD affects the over 204,000 medical students in Brazil. This study aims to explore the patterns of GBD experienced by medical students in Brazil. This is a cross-sectional study using an anonymous, Portuguese survey disseminated in June 2021 among Brazilian medical students. The survey was composed of 24 questions to collect data on GBD during medical school, formal methods for reporting GBD, and possible solutions for GBD. Of 953 responses, 748 (78%) were cisgender women, 194 (20%) were cisgender men, and 11 (1%) were from gender minorities. 65% (616/942) of respondents reported experiencing GBD during medical school. Women students experienced GBD more than men (77% versus 22%; P < 0.001). On comparing GBD perpetrator roles, both women (82%, 470/574) and men (64%, 27/42) reported the highest rate of GBD by faculty members. The occurrence of GBD by location differed between women and men. Only 12% (115/953) of respondents reported knowing their institution had a reporting mechanism for GBD. Most respondents experienced GBD during medical school. Cisgender women experienced GBD more than cisgender men. Perpetrators and location of GBD differed for men and women. Finally, an alarming majority of students did not know how to formally report GBD in their schools. It is imperative to adopt broad policy changes to diminish the rate of GBD and its a consequential burden on medical students.


Application of a dual channel peroneal nerve stimulator in a patient with a "central" drop foot.
Dropped foot is a common mobility problem amongst patients after a cerebro vascular accident. The condition arises from paresis of the muscles that control the foot movement during the swing phase of gait. If the abnormal movement is not compensated for, it results in a significant decrease in the mobility and hence quality of life. Compensation for the drop foot can be achieved through the application of functional electrical stimulation. To date, in the clinical environment, the stimulation has been applied through electrodes placed on the skin over the common peroneal nerve, and using a single channel implant device. It is well known that with these techniques it is difficult to establish a balanced response of the foot. An implantable dual channel system for stimulation of the deep and superficial peroneal nerve has now been developed for patients with a drop foot following a stroke. By stimulation of the two branches of the common peroneal nerve separately it is possible to achieve a precisely balanced dorsal flexion and eversion of the foot. Stimulation occurs via small bipolar electrodes which are placed subepineural. After successful tests on animals we have now started the two channel peroneal nerve stimulator implantation in patients. The preliminary results of the first implants are presented.


Morphological and rheological properties of cellulose nanofibrils prepared by post-fibrillation endoglucanase treatment.
Post-fibrillation endoglucanase treatment was employed to reduce the entanglement of cellulose nanofibrils (CNFs) produced with low extent of mechanical milling. The results showed that post-fibrillation endoglucanase treatment can effectively reduce the fibril degree of polymerization (DP) values and therefore entanglement. Cellulose DP was reduced from 520 for CNFs prepared by milling using a SuperMassColloider with 1 pass to 225 after endoglucanase treatment at a low dosage of 1 mg protein/g fibril for 24 h. Endoglucanase treatment can also effectively reduce the fibril mean diameter from 66 nm to 25 nm. Applying post-fibrillation endoglucanase treatment produced more uniform CNFs than the feed CNFs from pure mechanical treatment. All CNFs suspensions prepared by post-fibrillation endoglucanase treatment were non-Newtonian (except at very low concentrations below 0.5 wt%) with shear thinning behavior. Post-fibrillation endoglucanase treatment reduced the viscosity of CNF suspensions due to decreasing fibril DP values and the extent of entanglement.


Epilogue: a critical look at a nascent field.
A broad coalition of partners, entitled PHIRIC (Population Health Intervention Research Initiative for Canada), are working to increase the amount/quality of population health intervention research (PHIR) in our country. A central theme is to advance the science in this area. The current CJPH Supplement is welcomed as a set of diverse studies done to further our understanding of PHIR. The papers illustrate the range of questions that can be addressed and the variety of methods that need to be utilized. There is a need for critical reflection on three questions: 1) what constitutes PHIR? 2) which intervention parameters can be researched? and 3) what methods are recognized by the research community? Although legitimate attempts to define PHIR exist, the boundaries remain elusive. (Even the choice of articles in the current Supplement might be questioned.) It has cogently been argued that 'true' public health interventions intend to change risk conditions and alter distributions of health risk. This commentary suggests an important amendment to prior evaluation questions in stating that PHIR must pay attention to how intervention outcomes are distributed. There are also questions inherent in assessing the equity and distribution of an outcome. The bottom line is that we need to spread the word: more research is needed.


Angiopoietin-like protein 4 inhibition of lipoprotein lipase: evidence for reversible complex formation.
Elevated triglycerides are associated with an increased risk of cardiovascular disease, and lipoprotein lipase (LPL) is the rate-limiting enzyme for the hydrolysis of triglycerides from circulating lipoproteins. The N-terminal domain of angiopoietin-like protein 4 (ANGPTL4) inhibits LPL activity. ANGPTL4 was previously described as an unfolding molecular chaperone of LPL that catalytically converts active LPL dimers into inactive monomers. Our studies show that ANGPTL4 is more accurately described as a reversible, noncompetitive inhibitor of LPL. We find that inhibited LPL is in a complex with ANGPTL4, and upon dissociation, LPL regains lipase activity. Furthermore, we have generated a variant of ANGPTL4 that is dependent on divalent cations for its ability to inhibit LPL. We show that LPL inactivation by this regulatable variant of ANGPTL4 is fully reversible after treatment with a chelator.


Relationship between state medicaid policies, nursing home racial composition, and the risk of hospitalization for black and white residents.
To examine racial differences in the risk of hospitalization for nursing home (NH) residents. National NH Minimum Data Set, Medicare claims, and Online Survey Certification and Reporting data from 2000 were merged with independently collected Medicaid policy data. One hundred and fifty day follow-up of 516,082 long-stay residents. 18.5 percent of white and 24.1 percent of black residents were hospitalized. Residents in NHs with high concentrations of blacks had 20 percent higher odds (95 percent confidence interval [CI]=1.15-1.25) of hospitalization than residents in NHs with no blacks. Ten-dollar increments in Medicaid rates reduced the odds of hospitalization by 4 percent (95 percent CI=0.93-1.00) for white residents and 22 percent (95 percent CI=0.69-0.87) for black residents. Our findings illustrate the effect of contextual forces on racial disparities in NH care.


Detection of CMV DNA in clinical samples of AIDS patients by chemiluminescence hybridization.
A sensitive chemiluminescence dot-blot hybridization assay for the detection of CMV DNA in clinical samples of AIDS patients is described. In the chemiluminescence hybridization assay, digoxigenin-labelled CMV DNA probes were used and when hybridized they were detected by anti-digoxigenin Fab fragments conjugated with alkaline phosphatase. Adamantyl 1,2-dioxetane phenyl phosphate was used as the chemiluminescent substrate. The results were recorded by instant photographic films. The results obtained with the chemiluminescence hybridization assay were compared with the results obtained by hybridization with colourimetric detection. The chemiluminescent assay proved specific, sensitive and reliable and thus can be used as a valuable routine diagnostic test for the detection of CMV DNA in clinical samples.


[Torula thermophila, strain UzPT-1 a thermophilic producer of proteolytic enzymes].
The properties of the thermophilous fungus Torula thermophila producing proteolytic enzymes are described. Oprimal conditions for its cultivation are established. The maximum action of the preparation isolated from the culture liquid filtrate is at pH 9-11. The temperature maximum of the enzyme activity is 70 degrees C.


Critical care interventions in children aged 6 months to 12 years admitted to the pediatric intensive care unit after unintentional cannabis exposures.
Cannabis exposures in children have risen sharply in recent years, resulting in increased hospital visits and admission to pediatric intensive care units (PICUs). The intent of this study was to describe the proportion of pediatric patients admitted to the PICU after unintentional cannabis ingestion that received critical care interventions (CCIs) along with describing trends over time in hospitalization, admission to the PICU, and clinical effects and treatments outside of the PICU. This was a retrospective database study utilizing the National Poison Data System (NPDS) from 1/1/2000 to 12/31/2020. Children 6 months to 12 years of age with single substance cannabis exposures were included. A total of 12,882 cases were included. There was an increase in the proportion of cases seen in a hospital over time from 43.8% in 2000 to 54.6% in 2020 (range 29.1-62.6%). In patients seen in a HCF, the proportion admitted to the PICU was 9.5% in 2000 and 14% in 2020 (range: 5.6-29.0%). The 875 (6.8%) children admitted to the PICU were analyzed for the primary outcome. CCIs were performed in 69/875 (7.9%) cases that were admitted to the PICU. The most common CCIs in the PICU were intubation and sedation, 4.9 and 3.7%, respectively. Unintentional pediatric cannabis exposures are associated with clinically significant effects, including respiratory depression, hypotension, and bradycardia, but fewer than 5% of exposures were treated with CCIs, like intubation or vasopressors, in patients admitted to the PICU. Further work should assess specific reasons for admission to the PICU.


Structure of and kinetic channelling in bifunctional dihydrofolate reductase-thymidylate synthase.
The bifunctional enzyme dihydrofolate reductase-thymidylate synthase catalyses both the reductive methylation of 2'-deoxyuridylate and the subsequent reduction of dihydrofolate to yield 2'-deoxythymidylate and tetrahydrofolate at two spacially discrete sites situated on different protein domains. The X-ray structure of dihydrofolate reductase-thymidylate synthase from Leishmania major indicates that transfer of dihydrofolate between these sites does not occur by transient binding at both sites but rather by movement of dihydrofolate across the surface of the protein. The enzyme has an unusual surface charge distribution that could account for this channelling of dihydrofolate between active sites.


The relative importance of sources of greenhouse-gas emissions: comparison of global through subnational perspectives.
Increasing atmospheric concentrations of greenhouse gases are widely expected to cause global warming and other climatic changes. It is important to establish priorities for reducing greenhouse-gas emissions, so that resources can be allocated efficiently and effectively. This is a global problem, and it is possible, on a global scale, to identify those activities whose emissions have the greatest potential for enhancing the greenhouse effect. However, perspectives from smaller scales must be appreciated, because it is on scales down to the local level that response measures will be implemented. This paper analyzes the relative importance of emissions from the many individual sources, on scales ranging from global to national to subnational. Individual country perspectives and proposed policy measures and those of subnational political entities exhibit some commonalities but differ among themselves and from a global-scale perspective in detail.


Survival among motherless children in rural and urban areas in Guinea-Bissau.
With the increasing incidence of HIV and TB, motherless children are becoming a rapidly growing problem in Africa. However, few studies describe the survival patterns of these children. The aim of this study was to investigate the mortality of motherless children in urban and rural areas of Guinea-Bissau. A historical cohort study was set up in urban and rural areas in Guinea-Bissau. Motherless children were selected from two study cohorts under demographic surveillance since 1990. The relatives of 128 motherless children from the rural cohort and 192 from the urban area, as well as a total of 807 controls, were examined and interviewed. Controlling for significant background factors revealed that motherless children had a markedly higher mortality than that of controls in both urban (mortality rate ratio (MR) 2.32 (95% confidence interval 1.11-4.84)) and rural areas (MR = 4.16 (2.79-6.22)). Virtually all the excess mortality occurred among children under 2 y of age when their mother died. Few motherless children had been provided with surrogate breastfeeding. Since nearly all children in Guinea are breastfed until 2 y of age, premature weaning may be one of the major causes of the higher mortality rates observed among motherless children.


Low temperature cultivation of Escherichia coli carrying a rice lipoxygenase L-2 cDNA produces a soluble and active enzyme at a high level.
Using a T7 RNA polymerase promoter system, rice lipoxygenase L-2 cDNA was expressed in E. coli as a fusion protein with 18 amino acid residues at the amino terminal end of the original enzyme. Incubation at 37 degrees C for 3 h in the presence of the inducer resulted in the production of inactive lipoxygenase. However, when induction was carried out at 15 degrees C for 16 h, active lipoxygenase, amounting to 3% of the total soluble protein, was produced. The enzyme was purified by ammonium sulfate precipitation and Mono-Q column chromatography to homogeneity at a yield of 80%. Expression of this protein should permit future site-directed mutagenesis of the gene and crystallization of the enzyme.


Common mechanisms for declines in oxidative stress tolerance and proliferation with aging.
Aging is often characterized by reduced stress tolerance and decreased proliferative capacity, but little is known about the effects of aging on signaling pathways important in regulating these responses. Recent studies from our laboratory have implicated impairments in epidermal growth factor receptor (EGFR) signaling and extracellular signal-regulated kinase (ERK) activation to both effects in rat hepatocytes. Here we investigated the responsiveness of hepatocytes derived from young (4-5 months) and aged (24-29 months) mice to proliferative signals (low concentrations of H2O2 and epidermal growth factor [EGF] stimulation), and oxidant injury (high H2O2 concentrations). Old hepatocytes displayed lower levels of DNA synthesis in response to low H(2)O(2) concentrations (5-10 microM) and EGF stimulation, and reduced survival following treatment with high H2O2 concentrations (20-50 microM). Both effects were associated with reduced activation of ERK and diminished phosphorylation of EGFR tyrosine residue 1173. p38 was also activated by H2O2, but to a greater extent in old cells. Pharmacologic inhibition of ERK increased the sensitivity of young cells to H2O2-induced cell death, while inhibition of p38 decreased the sensitivity of old cells. Our findings suggest that impairments in common signaling events underlie age-related declines in proliferative capacity and oxidative stress tolerance in mouse hepatocytes, and that an imbalance in ERK and p38 activities contributes to the greater sensitivity of aged cells to H2O2.


Mutation of the NPM1 gene contributes to the development of donor cell-derived acute myeloid leukemia after unrelated cord blood transplantation for acute lymphoblastic leukemia.
Donor cell leukemia (DCL) is a rare but severe complication after allogeneic stem cell transplantation. Its true incidence is unknown because of a lack of correct recognition and reporting, although improvements in molecular analysis of donor-host chimerism are contributing to a better diagnosis of this complication. The mechanisms of leukemogenesis are unclear, and multiple factors can contribute to the development of DCL. In recent years, cord blood has emerged as an alternative source of hematopoietic progenitor cells, and at least 12 cases of DCL have been reported after unrelated cord blood transplantation. We report a new case of DCL after unrelated cord blood transplantation in a 44-year-old woman diagnosed as having acute lymphoblastic leukemia with t(1;19) that developed acute myeloid leukemia with normal karyotype and nucleophosmin (NPM1) mutation in donor cells. To our knowledge, this is the first report of NPM1 mutation contributing to DCL development.


Lissencephaly: computed tomographic diagnosis.
Lissencephaly is a rare congenital condition with distinctive brain changes. Most of these changes were demonstrated on computed tomography, and include lack of cortical sulci and gyri; calcification in the region of paraphysis; wide, shallow sylvian fissures; colpocephaly; poor development of white matter; and persistent cavum septum pellucidum and cavum vergae.


[Use of a method of regulated breathing to improve pulmonary respiratory function and to enhance the efficacy of aerosol therapy].
A new technique of controlled respiration is proposed which warrants longer exposure of the bronchi, bronchioles and alveoli in the lungs to chemotherapeutic drugs. These penetrate deeply into the target pulmonary tissue affected in lobar left- or right-side pneumonias, pulmonary atelectases. The technique promotes more efficient and less durable treatment in patients with respiratory and pulmonary disorders.


Characterisation of Isospora lusitanensis n. sp. (Apicomplexa: Eimeriidae) from the Eurasian blackbird Turdus merula Linnaeus (Passeriformes: Turdidae) in mainland Portugal.
A new coccidian species (Protozoa: Apicomplexa: Eimeriidae) is described from a specimen of the Eurasian blackbird Turdus merula Linnaeus held for rehabilitation and reintroduction into the wild in a centre for research and recovery of wild animals in Quinta de Marim, Olhão, Portugal. Isospora lusitanensis n. sp. has subspherical to ovoidal oöcysts, measuring on average 26.4 × 23.4 μm, with smooth, bi-layered wall c.1.1 μm thick. Micropyle and oöcyst residuum are absent, but a polar granule is present. Sporocysts are ellipsoidal, measuring on average 16.0 × 10.9 µm. Stieda body is knob-like and sub-Stieda body is prominent and rounded. Sporocyst residuum is composed of scattered spherules. Sporozoites are vermiform, with one refractile body and a nucleus. The morphological and morphometric data for the new species were compared with those for species parasitising birds of the Muscicapidae, Turdidae, Timaliidae, Troglodytidae and Cinclidae, which are considered phylogenetically close. The original histograms of Isospora turdi Schwalbach, 1959 were redrawn for comparison with I. lusitanensis n. sp. and a linear regression of width against length of the oöcysts is presented for characterisation. This is the first isosporoid coccidian described from T. merula in mainland Portugal.


Altered Cerebrospinal Fluid (CSF) in Children with Ataxia Telangiectasia.
Ataxia telangiectasia (A-T) is a devastating multi-system disorder characterized by progressive cerebellar ataxia and immunodeficiency. The neurological decline may be caused by multiple factors of which ongoing inflammation and oxidative stress may play a dominant role. The objective of the present investigation was to determine cerebrospinal fluid (CSF) proteins and possible low-grade inflammation and its relation to age and neurological deterioration. In the present study, we investigated 15 patients with A-T from 2 to 16 years. Our investigation included blood and CSF tests, clinical neurological examination, A-T score, and MRI findings. The albumin ratio (AR) was analyzed to determine the blood-brain-barrier function. In addition, inflammatory cytokines (IL-1α, IL-6, IL-8, IL-12 p40, IL-17A, IFN-γ, TNF-α) were measured by the multiplex cytometric bead array. We compared the results with those from an age-matched control group. Three of the A-T patients were analyzed separately (one after resection of a cerebral meningioma, one after radiation and chemotherapy due to leukemia, one after stem cell transplantation). Patient had significantly more moderate and severe side effects due to CSF puncture (vomiting, headache, need for anti-emetic drugs) compared with healthy controls. Total protein, albumin, and the AR increased with age indicating a disturbed blood barrier function in older children. There were no differences for cytokines in serum and CSF with the exception of IL-2, which was significantly higher in controls in serum. The AR is significantly altered in A-T patients, but low-grade inflammation is not detectable in serum and CSF.


Effect of cornea material stiffness on measured intraocular pressure.
Intraocular pressure (IOP) in the human eye as measured by a Goldmann applanation tonometer (GAT) is known to be affected by individual differences in central corneal thickness (CCT). However, data from clinical studies also show considerable scatter in the correlation between measured IOP and CCT. One possible implication of the large observed scatter is that the true IOP (IOPT) also depends significantly on individual variations in the material stiffness properties of the cornea. This hypothesis is explored and evaluated herein using computational simulation of applanation tonometry. A simplified 2D finite element model of the eye, which employs a calibrated nonlinear transversely isotropic material model for the cornea, is developed, and a series of GAT simulations is carried out to study the effect of geometry and material properties of the cornea on the IOP readings obtained via GAT. The results of this parametric study provide a simple correction equation, which quantifies the effect on measured IOP of variations in CCT and corneal material stiffness. In addition, several previously proposed IOP correction equations are compared with the one proposed here.


Motor axon sprouting in frog sartorius muscles is not altered by contralateral axotomy.
Sartorius muscles of the frog Rana pipiens were used to study the incidence of motor nerve sprouting in normal unoperated muscles, in experimental muscles contralateral to axotomy of the sartorius nerve, and in sham-operated control muscles. Muscles were stained with either a combination of nitroblue tetrazolium nerve terminal stain and cholinesterase stain or with a combination of silver nerve terminal stain and cholinesterase stain. Each endplate that could be clearly seen was classified into one or more of the following categories: normal endplates without sprouts, three types of terminal sprouts, preterminal sprouts, nodal sprouts, sprouts of unknown origin and destination, and doubly innervated gutters. A quantitative study of 318 endplates from nine unoperated muscles, 779 endplates from 45 experimental muscles, and 694 endplates from 41 control muscles showed that all muscles had a high incidence of motor nerve sprouting and other forms of remodelling (20-28% of all endplates). There were, however, no significant differences between experimental, control, and unoperated muscles when results obtained with the same stains were compared. Results obtained with the two different stains were only slightly different. We conclude that sprouting is a very common but highly variable feature of normal frog neuromuscular junctions, and in the sartorius, contralateral axotomy does not alter this ongoing remodelling.


Thiazide Use and Cardiovascular Events in Type 2 Diabetic Patients With Well-Controlled Blood Pressure.
Evidence regarding the efficacy and safety of thiazides in patients with well-controlled and relatively low blood pressure (BP) is lacking. This study aimed to assess whether thiazide use is effective and safe in type 2 diabetic patients with well-controlled BP and whether intensive BP control leads to decreased risk of cardiovascular events depending on thiazide use. We performed an observational cohort study using data from the ACCORD study (Action to Control Cardiovascular Risk in Diabetes). The primary outcome was major adverse cardiovascular events (MACE), which was a composite end point including cardiovascular death, myocardial infarction, and stroke. Hazard ratios for primary and secondary outcomes with 95% CIs were calculated using Cox proportional hazards models. We included 10 011 type 2 diabetic patients. The overall mean follow-up period was 7.7 years, and 1776 patients experienced MACE. Mean systolic BP at baseline in patients taking and not taking thiazides was 137.2 and 135.7 mm Hg, respectively. Thiazide use was associated with increased risk of MACE, particularly stroke (hazard ratio, 1.49 [95% CI, 1.18-1.88]). In addition, thiazide use was significantly associated with higher risks of MACE and stroke in patients receiving intensive BP control but not in those receiving standard BP control. Similar associations were observed in analyses using propensity score matching. Intensive BP control reduced the risks of MACE and stroke in patients not taking thiazides but not in patients taking thiazides. Thiazide use may be harmful in type 2 diabetic patients with relatively low BP.


Effect of low temperature and relative humidity on reproduction and survival of the tick Rhipicephalus microplus.
The objective of this study was to evaluate the survival of Rhipicephalus microplus females under low temperature, to determine the influence of temperature and relative humidity (RH) on the biological parameters of this tick. In total 300 viable engorged female ticks collected from cattle were used. On the day of collection (D0), 30 female ticks were divided into three groups: G1, kept at 28 °C and 80% RH; G2, kept at 20 °C and 80% RH; and G3, kept at 20 °C and 30% RH. The remaining females were stored in a refrigerator at 4 °C. Over the next 9 days (D+1 to D+9), 30 female ticks were removed daily from the refrigerator and distributed among the three treatments. Egg mass, feed conversion rate, egg incubation period, larval hatch rate, number of dead females per group, weight of female tick removed from the refrigerator, and female tick weight loss were recorded. At 20 °C and 80% RH hatching was delayed. Among the female ticks kept in the refrigerator and then allocated to groups G1, G2, and G3, there was a reduction in reproductive efficiency and a rise in mortality as time in the refrigerator increased. The female ticks expressed their best reproductive capacity when subjected to the refrigeration temperature for a maximum of 3 days and then kept at 28 °C and 80% RH. Thus, although female ticks can be stored in the refrigerator before being sent to research centers for use in bioassays, the storage time should be as short as possible, not exceeding 3 days.


Prioritizing single-nucleotide variations that potentially regulate alternative splicing.
Recent evidence suggests that many complex diseases are caused by genetic variations that play regulatory roles in controlling gene expression. Most genetic studies focus on nonsynonymous variations that can alter the amino acid composition of a protein and are therefore believed to have the highest impact on phenotype. Synonymous variations, however, can also play important roles in disease pathogenesis by regulating pre-mRNA processing and translational control. In this study, we systematically survey the effects of single-nucleotide variations (SNVs) on binding affinity of RNA-binding proteins (RBPs). Among the 10,113 synonymous SNVs identified in 697 individuals in the 1,000 Genomes Project and distributed by Genetic Analysis Workshop 17 (GAW17), we identified 182 variations located in alternatively spliced exons that can significantly change the binding affinity of nine RBPs whose binding preferences on 7-mer RNA sequences were previously reported. We found that the minor allele frequencies of these variations are similar to those of nonsynonymous SNVs, suggesting that they are in fact functional. We propose a workflow to identify phenotype-associated regulatory SNVs that might affect alternative splicing from exome-sequencing-derived genetic variations. Based on the affecting SNVs on the quantitative traits simulated in GAW17, we further identified two and four functional SNVs that are predicted to be involved in alternative splicing regulation in traits Q1 and Q2, respectively.


Rapid analysis of Callicarpa L. using direct spray ionization mass spectrometry.
Direct spray such as leaf spray and paper spray ionization mass spectrometry (MS) is a powerful type of ambient MS for phytochemical analysis. In this paper, direct spray MS methods to rapidly distinguish and analyze five species of Callicarpa L. have been developed. To distinguish species, leaf spray MS was employed to directly analyze leaves. A small triangular leaf sample was wetted with 15 μL of spray solvent and a high DC voltage was then simply applied to the wet leaf sample, which was positioned in front of the inlet of a mass spectrometer to produce electrospray ionization. The MS signals of phenylpropanoid glycosides, i.e. forsythiaside B, poliumoside, verbascoside in leaves could be sensitively detected. The content characteristics of the phenylpropanoid glycosides in five species including Callicarpae kwangtungensis Folium, Callicarpae macrophyllae Folium, Callicarpa nudiflora Folium, Callicarpae formosanae Folium, Callicarpa longissima Folium could be used to distinguish them, then the mass spectra of the Callicarpa L. samples were analyzed using principal component analysis(PCA) or partial least squares-discriminant analysis(PLS-DA). For the rapid semi-quantitative analysis, of phenylpropanoid glycosides in leaves, paper spray MS was employed to determine phenylpropanoid glycosides in the extracts of Callicarpa L. leaves. Ginsenoside Rg1 was selected as an internal standard (I.S.). The calibration curves were constructed through ratios of target ion abundance to I.S. ion abundance vs. concentration of targets. The linearity range was 8-250 μg/mL (R(2)=0.9947) for forsythiaside B, 9-280 μg/mL (R(2)=0.9939) for verbascoside, and 9-260 μg/mL (R(2)=0.9917) for poliumoside, respectively. The limit of detection (LOD) was 1 μg/mL, 0.5 μg/mL and 1 μg/mL for forsythiaside B, verbascoside, and poliumoside, respectively.


Endoscopic resection of a large Brunner's gland hamartoma.
Brunner's Gland Hamartoma (BGH) is a benign tumor of the duodenum that can lead to gastrointestinal bleeding and intestinal obstruction. Endoscopic resection has seldom been reported. We describe the case of a duodenal obstruction caused by a large BGH (6 cm x 4 cm). We report a 57-year-old woman hospitalized for tarry stools, weight loss and anorexia. Endoscopy revealed a large BGH (6 cm x 4 cm). Endoscopic ultrasound (EUS) revealed a submucosal duodenal tumor. In this paper, we report a case of large hyperplasia of BGH, successfully treated by endoscopic technique.


Reduction of alloantibodies via proteasome inhibition in cardiac transplantation.
The presence of alloantibodies in patients awaiting heart transplantation is associated with increased waiting time to transplant, increased risk of rejection after transplant, an increased risk of cardiac allograft vasculopathy, and decreased survival. So far, treatments to reduce circulating antibodies to allow transplantation have been limited. We report the first clinical experience using a plasma-cell-depleting strategy with bortezomib to reduce anti-HLA antibodies in the heart transplant population. Six patients awaiting cardiac transplantation demonstrated persistently elevated anti-HLA antibodies, despite receiving a course of treatment with intravenous immunoglobulin (IVIg) and rituximab. These patients then underwent supplemental therapy with bortezomib in conjunction with plasmapheresis. One additional patient awaiting cardiac transplantation with elevated anti-HLA antibodies required bortezomib treatment for amyloidosis. Antibody strength was monitored after completion of treatment by solid-phase (single-antigen-bead) assay. The mean calculated panel-reactive antibody (cPRA) was reduced from 62% to 35% following a course of bortezomib (p = 0.01). Six of 7 patients demonstrated a significant decline in antibody levels. One patient remained refractory to desensitization therapy and died from sepsis while awaiting heart transplantation. Four patients successfully underwent cardiac transplantation without evidence of rejection or graft dysfunction. One patient developed early post-transplant graft dysfunction and died at 1 month from sepsis. Infection was the most common adverse effect associated with desensitization. In this pilot study, use of plasmapheresis and bortezomib appeared to decrease cPRA, even in patients refractory to desensitization with IVIg/rituximab, thus increasing the chances that an acceptable donor heart will be available for the sensitized patient awaiting heart transplantation. However, desensitization is associated with an increased risk of infection. Further studies are warranted to determine whether the benefits of desensitization using this strategy outweigh the risks.


Regulation of MCL1 through a serum response factor/Elk-1-mediated mechanism links expression of a viability-promoting member of the BCL2 family to the induction of hematopoietic cell differentiation.
Proliferation, differentiation, and apoptosis are tightly regulated during hematopoiesis, allowing amplification along specific lineages while preventing excessive proliferation of immature cells. The MCL1 member of the BCL2 family is up-regulated during the induction of monocytic differentiation (approximately 10-fold with 12-O-tetradecanoylphorbol 13-acetate (TPA)). MCL1 has effects similar to those of BCL2, up-regulation promoting viability, but differs from BCL2 in its rapid inducibility and its pattern of expression. Nuclear factors that regulate MCL1 transcription have now been identified, extending the previous demonstration of signal transduction through mitogen-activated protein kinase. A 162-base pair segment of the human MCL1 5'-flank was found to direct luciferase reporter activity, allowing approximately 10-fold induction with TPA that was suppressible upon inhibition of the extracellular signal-regulated kinase (ERK) pathway. Serum response factor (SRF), Elk-1, and Sp1 bound to cognate sites within this segment, SRF and Elk-1 acting coordinately to affect both basal activity and TPA inducibility, whereas Sp1 affected basal activity only. Thus, the mechanism of the TPA-induced increase in MCL1 expression seen in myelomonocytic cells at early stages of differentiation involves signal transduction through ERKs and transcriptional activation through SRF/Elk-1. This finding provides a parallel to early response genes (e.g. c-FOS and EGR1) that affect maturation commitment in these cells and therefore suggests a means through which enhancement of cell viability may be linked to the induction of differentiation.


Can music therapy and aroma therapy really reduce dental anxiety and fear?
This study was a randomised controlled trial (RCT) designed to evaluate the effectiveness of music therapy and aromatherapy on reducing dental anxiety and fear in children during a fissure sealant treatment. Participants were selected from three primary schools from the Phitsanulok Province in Thailand. These children were randomly allocated to a (1) Control group (n = 33), (2) Music Therapy Experimental Group (n = 33), (3) Aromatherapy Group (n = 33) and (4) Combination Experimental Group (n = 33). Measures included dental anxiety and fear scores as well as vital signs (blood pressure, heart rate and oxygen saturation level) pre- and post-fissure sealant treatment. Descriptive statistics were used for categorical data. Mean and standard deviations were calculated for continuous data. For the four arms, Chi-squared tests and one-way ANOVA were used to compare base line characteristics of the participants. Paired t-tests were used to compare changes within groups in dental anxiety and fear and vital signs pre- and post-treatment. Overall, 128 participants took part in were in the final analysis (32, 33, 31 and 32 in groups 1-4, respectively). In the music therapy group and aromatherapy group, there was a decrease in dental anxiety and fear, blood pressure and increase in oxygen saturation level (p = 0.05). In the combination group, there was a decrease in dental anxiety and fear, heart rate, blood pressure and increase in oxygen saturation level (p = 0.05). A statistically significant interaction effect was found between combination therapy on dental anxiety and fear (F = 22.22; P < 0.001) and oxygen saturation level (F = 17.40; P < 0.001). The authors conclude that the findings of this RCT suggest that the use of music therapy and aromatherapy reduces dental anxiety and fear in children, and is more effective when used in combination.


Chondroitin sulfate-based nanoparticles for enhanced chemo-photodynamic therapy overcoming multidrug resistance and lung metastasis of breast cancer.
As a major therapeutic approach for cancer treatment, the effectiveness of chemotherapy is challenged by multidrug resistance (MDR). Herein, we fabricated novel redox-responsive, chondroitin sulfate-based nanoparticles that could simultaneously deliver quercetin (chemosensitizer), chlorin e6 (photosensitizer) and paclitaxel (chemotherapeutic agent) to exert enhanced chemo-photodynamic therapy for overcoming MDR and lung metastasis of breast cancer. In vitro cell study showed that nanoparticles down-regulated the expression of P-glycolprotein (P-gp) on MCF-7/ADR cells and thereby improved the anticancer efficacy of PTX against MCF-7/ADR cells. Moreover, NIR laser irradiation could induce nanoparticles to generate cellular reactive oxygen species (ROS), leading to mitochondrial membrane potential loss, and meanwhile facilitating lysosomal escape of drugs. Importantly, the novel nanoplatform exhibited effective in vivo MDR inhibition and anti-metastasis efficacy through enhanced chemo-photodynamic therapy. Thus, the study suggested that the multifunctional nanoplatform had good application prospect for effective breast cancer therapy.


Local-to-global form interference in biological motion perception.
Point-light walkers have been useful to study the contribution of form and motion to biological motion perception by manipulating the lifetime, number, or spatial distribution of the light points. Recent studies have also manipulated the light points themselves, replacing them with small images of objects. This manipulation degraded the recognizability of biological motion, particularly for local images of human bodies. This result suggests an interference of body form information in the local images with the body form analysis necessary for global biological motion recognition at the global level. We further explored this interference with respect to its selectivity for body orientation and motion. Participants had to either discriminate the facing direction (left/right) or the walking direction (forward/backward) of a global walker composed of local stick figures that could face left or right and either stand still or walk forward or backward. Local stick figures interfered stronger with the facing direction task if they were facing in the same direction as the global walker. Walking (forward/backward/static) of the stick figures influenced neither the facing direction task nor the walking direction task. We conclude that the interference is highly specific since it concerns not only the category (human form), but even the facing direction.


Biosorption of Cr (VI) by acid-modified based-waste fungal biomass from Calocybe indica fruiting bodies production.
The world is going through a colossal drinking water scarcity. Unchecked discharge (even at trace levels) of Cr (VI) from industries into water bodies is a serious environmental concern. Here, we report waste fungal biomass (WFB) for the detoxification and removal of chromium ions. Biomass understudy was collected from Calocybe indica fruiting bodies. WFB was used after drying and pretreatment with two distinctive chemical methods, which improved the remediation effectiveness of Cr (VI). Light microscope and Field emission Scanning microscope (FESEM) were employed to elucidate the surface morphology of waste fungal biomass. While Fourier-Transform Infrared-Spectroscopy (FTIR) and Energy Dispersive X-Ray analysis (EDAX) were deployed to explore the mechanism of interaction between Cr (VI) anion and waste fungal biomass. X-ray Photoelectron Spectroscopy (XPS) analyses demonstrated considerable conversion of Cr (VI) into nontoxic Cr (III) species. The most favorable condition for optimum Cr (VI) remediation of 99.66% by treated waste fungal biomass (TWFB) occurred at pH 3, contact time 10 min, adsorbent dosage 3 gL-1, Cr (VI) concentration 4 mgL-1, stirring speed 140 rpm, and temperature 320 K, where for untreated waste fungal biomass (UWFB) the optimum of 85% remediation occurred at a contact time 15 min, and adsorbent dosage 2 gL-1 whereas other experimental conditions remained identical as TWFB biosorbent. Pseudo-second-order kinetics (R2 > 0.99) model matched the adsorption rate. And, the Freundlich isotherm model (R2 > 0.99) is shown to be a better match for the experimental data. The optimum amount of Cr (VI) adsorbed by the TWFB and UWFB were 205.8 ± 10.1 and 72.85 ± 2.36 mgg-1, respectively. Thermodynamic parameters revealed that the adsorption was spontaneous (ΔG ˂ 0), endothermic (ΔH > 0), and entropy-driven (ΔS > 0). The generated WFB adsorbent also has significant recycling potential. After five cycles of regeneration and adsorption. It can still keep up good remediation effectiveness of Cr (VI) ions to 85.5.


Uptake of Heavy Metals from Industrial Wastewater Using In Vitro Plant Cultures.
The plant species Bacopa monnieri has been observed to reduce the heavy metal concentrations in its vicinity. The present study is a comparison of in vitro culture and soil-grown plants of B. monnieri to remove Cr and Cd, from synthetic solution and effluent obtained from industrial area. Results were obtained at every half hour interval upto 180 min. Samples were observed for light absorption using UV-Visible spectrophotometer. Statistically, both systems reclaimed Cr and Cd from polluted water. In vitro cultures showed 67% and 93% removal of Cr and Cd from industrial wastewater whereas soil-grown plants showed 64% and 83% Cr and Cd removal. However, reduction rate was significantly higher for in vitro culture as compared to soil-grown plants. Besides other advantages, in vitro plant cultures proved to be more potent to detoxify pollutants in less time. This approach can be used for the removal of heavy metals at large scale.


Unlinked regulation of cell growth and differentiation in immature B cell lines.
We established many immunoglobulin-null immature B cell lines transformed by tsOS-59, a temperature-sensitive mutant of Abelson murine leukemia virus. In different cell lines cell growth was depressed and cell differentiation (generation of intracytoplasmic mu-positive cells from Ig- cells) was induced by the shift of culture temperature from low (35 degrees C) to high (39 degrees C). Cell lines were categorized into four groups: (i) temperature sensitive (ts) to both cell growth and differentiation, (ii) ts to cell growth but not to cell differentiation, (iii) ts to cell differentiation but not to cell growth, and (iv) ts to neither cell growth nor differentiation. These results indicated that the depression of cell growth did not necessarily induce cell differentiation, and that cell differentiation was induced regardless of whether cell growth was depressed or not. Furthermore, the results showed that the depression of cell growth and the induction of cell differentiation occurred without the reduction of tyrosine kinase activity of P120gag-abl at high, nonpermissive temperature. Our cell growth and differentiation system described here should provide us with the interesting findings of the relation between B cell growth and differentiation.


Viscosity and flow properties of concentrated solutions of chitosan with different degrees of deacetylation.
The effects of the degree of deacetylation (DD) on the viscosity and flow behaviour of concentrated solutions of chitosan were investigated using 0.2 M CH3COOH and 0.2 M CH3COOH/0.1 M CH3COONa aqueous solutions as solvents. The results indicated that the viscosity and flow properties of the solutions differed with the DD of chitosan. The solution viscosities and the non-Newtonian flow properties as well as the flow activation energies E gamma increased with the increasing DD of chitosan. However, the additional salt decreased the viscosities and the non-Newtonian flow properties of the solutions of chitosan, but did not change the flow activation energies E gamma of the solutions.


[Assesssment of posttraumatic stress disorder of survivors in the "5. 12" earthquake after 3 years].
To investigate the posttraumatic symptom of survivors in the "5.12" earthquake and its influencing factors. Six hundred and five earthquake survivors from Beichuan District were interviewed by systematic sampling method. Posttraumatic symptoms and demographic source were measured using the PTSD Checklist Civilian Version (PCL-C) and self-questionnaire. The prevalence of PTSD is 11.2%. Survivors who suffer from PTSD commonly behave re-experiencing trauma and hyper arousal. The logistic regression shows that victims who have lost family members, who are females, and who suffered huge losses of property in this earthquake are the dangerous factors of PTSD. The results of OR are 2.848, 1.778 and 3.297 respectively. Posttraumatic symptoms are still common among victims three years after "5.12" Earthquake. Some efficient measures regarding the different survivors should be taken. Victims who have lost family members, who are females, and who suffered huge losses of property, should be paid more attention. Re-experiencing trauma and hyper arousal are symptoms that deserve our main concern.


The ethics of letting go.
Emphasis in medical training is laid on therapeutic approaches that prolong life at all costs. This training is reinforced by current medical practices in the United States and by an ever-expanding medical technology, such that it becomes difficult, if not impossible, to "let the patient go." In response, individuals promote the "right-to-die" and euthanasia, so that patients may maintain their human dignity during the dying process. A more rational and nonpolarized approach to this issue would be to strive for the preservation of human function, not physiological function alone. The goal would include helping patients advance their life plans and values. On this basis, therapies can be withheld and withdrawn when they no longer reflect these goals. Caution regarding potential abuses of this approach need to be raised, however, when considering care for the very vulnerable of society. In that regard, therapeutic goals directed at maintaining human function need to be developed in parallel with global societal values regarding quality of life issues.


The Chubut River estuary as a source of microplastics and other anthropogenic particles into the Southwestern Atlantic Ocean.
This is the first report of anthropogenic particles (APs), including microplastics and synthetic, semi-synthetic and anthropogenically-altered natural fibers, in water and sediment of the Chubut River estuary. This river is the main source of freshwater in Chubut Province (Patagonia, Argentina), where wastes and pollutants are poured and finally end in the Atlantic Ocean. The average concentration in surface and bottom water samples was 5.5 items/L, while in sediment was 175.4 items/kg dw. Raman's analysis identified particles dominated by polyethylene terephthalate (PET) (35.5 %), dye signature only (18.5) and anthropogenic cellulose (10 %). Fibers were the prevalent shape (83 %), and the chemical identification evidenced a textile origin. The highest APs concentration was found in sediments from the site with the finest grain size and the greatest amount of organic matter. Present results will provide a baseline for future studies and raise public and governmental awareness.


Some legal decisions significant for ophthalmology.
The court decisions included in this article reflect judicial attitudes towards medicolegal cases in most states. The courts have defined and amplified issues of informed consent, the physician's duty to warn patients and protect third parties, the limitations of the "Good Samaritan" statute, the responsibility of the physician to prepare insurance reports, and the denial of coverage by third party payers. Several cases regarding these issues are reviewed and their implications for ophthalmologists are discussed.


Status epilepticus as an initial presentation of Brugada syndrome: a case report.
Patients presenting with convulsions are sometimes incorrectly treated for epilepsy, as these symptoms may be manifestations of underlying cardiac disease. Brugada syndrome, associated with an elevated risk of developing fatal arrhythmic events, is a rare disorder characterized by a unique electrocardiographic pattern. The typical clinical presentation mainly involves syncope of unclear cause or sudden death. Seizures are uncommon clinical manifestations of Brugada syndrome, and reports of status epilepticus as initial presentation of Brugada syndrome are extremely rare. In this case report, we present a male patient with a typical pattern on 12-lead electrocardiography, following resuscitation for generalized convulsive status epilepticus.


Liquid/air partition coefficients of the trimethylbenzenes.
Liquid/air partition coefficients (lambda) of the three isomers of trimethylbenzene (1,2,3-,1,2,4-, and 1,3,5-TMB) were determined in vitro using automated head space gas chromatography. The liquids used were water (physiological saline), fresh human blood, and olive oil. The lambda water/air were 1.23 (95% confidence interval 1.11-1.35) for 1,3,5-TMB, 1.61 (1.47-1.75) for 1,2,4-TMB, and 2.73 (2.54-2.92) for 1,2,3-TMB. Corresponding lambda blood/air were 43.0 (40.8-45.2), 59.1 (56.9-61.3), and 66.5 (63.7-69.3). As expected, the TMBs have a high affinity for olive oil, the lambda oil/air being 9,880 (9,620-10,140), 10,200 (9,900-10,400), and 10,900 (10,500-11,300), respectively. It is notable that all partition coefficients increase in the order 1,3,5-TMB < 1,2,4-TMB < 1,2,3-TMB.


Anti-atherosclerotic effect of simvastatin depends on the presence of apolipoprotein E.
Low density lipoprotein receptor deficient (LDLR-KO) and apolipoprotein E deficient (apo E-KO) mice both develop hyperlipidemia and atherosclerosis by different mechanisms. The aim of the present study was to compare the effects of simvastatin on cholesterol levels, endothelial dysfunction, and aortic lesions in these two models of experimental atherosclerosis. Male LDLR-KO mice fed a high cholesterol (HC; 1%) diet developed atherosclerosis at 8 months of age with hypercholesterolemia. The addition of simvastatin (300 mg/kg daily) to the HC diet for 2 more months lowered total cholesterol levels by approximately 57% and reduced aortic plaque area by approximately 15% compared with the LDLR-KO mice continued on HC diet alone, P<0.05. Simvastatin treatment also improved acetylcholine (ACh)-induced endothelium-dependent vasorelaxation in isolated aortic rings, which was associated with an increase in NOS-3 expression by approximately 88% in the aorta measured by real time polymerase chain reaction (PCR), P<0.05. In contrast, in age-matched male apo E-KO mice fed a normal diet, the same treatment of simvastatin elevated serum total cholesterol by approximately 35%, increased aortic plaque area by approximately 15%, and had no effect on endothelial function. These results suggest that the therapeutic effects of simvastatin may depend on the presence of a functional apolipoprotein E.


Deconstructing the germinal center, one cell at a time.
Successful vaccination relies on driving the immune response towards high specificity, affinity and longevity. Germinal centers facilitate the evolution of antigen-specific B cells by iterative rounds of diversification, selection, and differentiation to memory and plasma cells. Experimental evidence points to B cell receptor affinity and amount of antigen presented to follicular helper T cells as main drivers of clonal evolution. Concurrent studies suggest that modifiers of cognate contact, temporal mechanisms, and stochastic factors can also shape diversity and influence differentiation to memory and plasma cells, but molecular pathways driving these selection decisions are unresolved. Due to rapid cycles of transcriptional change in the germinal center, single-cell resolution is imperative to dissect mechanisms dictating the mature antigen-specific repertoire. Future studies linking high-resolution analysis of this diverse evolving population with cellular outcome are needed to fully understand the complex mechanisms of selection driving antigen-specific humoral immunity.


Autoimmune neutropenia and thrombocytopenia associated with development of antibodies to human immunodeficiency virus.
We report on two patients with acute human immunodeficiency virus (HIV) infection, who developed an infectious mononucleosis-like clinical episode with thrombocytopenia and granulocytopenia. In both cases we detected the presence of IgG antigranulocyte antibodies and in one case the presence of IgG, IgM and IgA antiplatelet antibodies. The mechanisms of these cytopenias are discussed. The association between such autoimmune cytopenias and acute HIV infection has not been previously reported. We suggest testing for HIV seroconversion in patients with a seronegative infectious mononucleosis-like syndrome belonging to groups with a high risk for HIV infection.


Structure of the human transferrin receptor-transferrin complex.
Iron, insoluble as free Fe(3+) and toxic as free Fe(2+), is distributed through the body as Fe(3+) bound to transferrin (Tf) for delivery to cells by endocytosis of its complex with transferrin receptor (TfR). Although much is understood of the transferrin endocytotic cycle, little has been uncovered of the molecular details underlying the formation of the receptor-transferrin complex. Using cryo-electron microscopy, we have produced a density map of the TfR-Tf complex at subnanometer resolution. An atomic model, obtained by fitting crystal structures of diferric Tf and the receptor ectodomain into the map, shows that the Tf N-lobe is sandwiched between the membrane and the TfR ectodomain and that the C-lobe abuts the receptor helical domain. When Tf binds receptor, its N-lobe moves by about 9 A with respect to its C-lobe. The structure of TfR-Tf complex helps account for known differences in the iron-release properties of free and receptor bound Tf.


Integrative computational approach for genome-based study of microbial lipid-degrading enzymes.
Lipid-degrading or lipolytic enzymes have gained enormous attention in academic and industrial sectors. Several efforts are underway to discover new lipase enzymes from a variety of microorganisms with particular catalytic properties to be used for extensive applications. In addition, various tools and strategies have been implemented to unravel the functional relevance of the versatile lipid-degrading enzymes for special purposes. This review highlights the study of microbial lipid-degrading enzymes through an integrative computational approach. The identification of putative lipase genes from microbial genomes and metagenomic libraries using homology-based mining is discussed, with an emphasis on sequence analysis of conserved motifs and enzyme topology. Molecular modelling of three-dimensional structure on the basis of sequence similarity is shown to be a potential approach for exploring the structural and functional relationships of candidate lipase enzymes. The perspectives on a discriminative framework of cutting-edge tools and technologies, including bioinformatics, computational biology, functional genomics and functional proteomics, intended to facilitate rapid progress in understanding lipolysis mechanism and to discover novel lipid-degrading enzymes of microorganisms are discussed.


Psychotherapeutic issues encountered in the psychotherapy of chronic pain patients.
The clinical art of applying scientific knowledge to a patient's circumstances of suffering is especially challenging when practicing psychotherapy with people who are experiencing chronic pain. Health psychology has made substantial contributions by showing that psychosocial factors are essential in explaining the phenomenon of pain and, particularly, its chronic progression to debilitating degrees. In addition, clinical health psychology has developed empirically supported treatments that improve the lives of chronic pain sufferers across many dimensions. This article is not concerned with reviewing that evidence or the treatment programs, but with noting some issues pain management psychotherapists may face in using this knowledge effectively to improve the lives of their patients.


Isoforms of NO-sensitive guanylyl cyclase.
By the formation of cGMP the NO-sensitive guanylyl cyclase plays a key role within the NO/cGMP signaling cascade involved in vascular regulation and neurotransmission. The prosthetic heme group of the enzyme acts as the NO sensor, and binding of NO induces conformational changes leading to an up to 200-fold activation of the enzyme. The unexpected fast dissociation half-life of NO of a few seconds is fast enough to account for the deactivation of the enzyme in biological systems. YC-1 and its analogues acting as NO sensitizers uncovered a new pharmacologically and conceivably physiologically relevant regulatory principle of the enzyme. Two existing isoforms of the heterodimeric guanylyl cyclase (alpha1beta1, alpha2beta1) are known that are functionally indistinguishable. Up to now, the NO-sensitive guanylyl cyclase has been considered as a soluble enzyme. However, recent evidence about the alpha2beta1 isoform interacting with a PDZ domain of the postsynaptic scaffold protein PSD-95 suggests that the alpha2 subunit directs a membrane association of this isoform. The interaction with PSD-95 locates the alpha2beta1 isoform in close proximity to the NO-generating NO synthase thereby enabling the NO sensor to respond to locally raised NO concentrations.


Down-regulation of USP13 mediates phenotype transformation of fibroblasts in idiopathic pulmonary fibrosis.
Idiopathic pulmonary fibrosis (IPF) is a fatal disease characterized by fibroblastic foci and progressive scarring of the pulmonary parenchyma. IPF fibroblasts display increased proliferation and enhanced migration and invasion, analogous to cancer cells. This transformation-like phenotype of fibroblasts plays an important role in the development of pulmonary fibrosis, but the mechanism for this is not well understood. In this study, we compared gene expression profiles in fibrotic lung tissues from IPF patients and normal lung tissues from patients with primary spontaneous pneumothorax using a cDNA microarray to examine the mechanisms involved in the pathogenesis of IPF. In a cDNA microarray, we found that USP13 was decreased in lung tissues from patients with IPF, which was further confirmed by results from immunohistochemistry and western blot assays. Then, we used RNA interference in MRC-5 cells to inhibit USP13 and evaluated its effects by western blot, real-time RT-PCR, bromodeoxyuridine incorporation, and transwell assays. We also used co-immunoprecipitation and immunofluorescence staining to identify the correlation between USP13 and PTEN in IPF. USP13 expression levels were markedly reduced in fibroblastic foci and primary IPF fibroblast lines. The depletion of USP13 resulted in the transformation of fibroblasts into an aggressive phenotype with enhanced proliferative, migratory, and invasive capacities. Additionally, USP13 interacted with PTEN and mediated PTEN ubiquitination and degradation in lung fibroblasts. Down-regulation of USP13 mediates PTEN protein loss and fibroblast phenotypic change, and thereby plays a crucial role in IPF pathogenesis.


[Development of medicine in Lwów (Lemberg, Lviv). Part 1: beginnings and establishment of urology until 1944].
Innovations in diagnostic techniques due to the introduction of endoscopy and the development of X‑ray technology were fundamental in reducing the dependence on surgery and to recognize urology as a new discipline. Thereafter, endoscopic surgery came to the fore. The aim of this article is to present the development of urology in Lviv as an independent discipline and its distinction from surgery. Well-known Lviv surgeons who were interested in surgery of the urogenital system are discussed. The beginnings of urology and its development within the framework of surgical departments and ultimately as an independent institution in the interwar period are described. The fate of the Polish department of urology in the interwar period and during the Second World War is also described. J. Molendziński, G. Ziembicki, Z. Leńko and S. Laskownicki can be described as the founders of urology in Lviv (Lemberg). Extensive research in archives and libraries in Poland and Ukraine has been undertaken in the preparation of this article.


A CRISPR view of gene regulation.
Due to plummeting costs, whole genome sequencing of patients and cancers will soon become routine medical practice; however, we cannot currently predict how non-coding genotype affects cellular gene expression. Gene regulation research has recently been dominated by observational approaches that correlate chromatin state with regulatory function. These approaches are limited to the available genotypes and cannot scratch the surface of possible sequence combinations, and thus there is a need for perturbation-based approaches to better understand how DNA encodes gene regulatory functions. CRISPR/Cas9 genome editing has revolutionized our ability to alter genome sequence, and CRISPR/Cas9-based assays have already begun to contribute to new paradigms of gene regulation. We discuss the variety of arenas in which current and future CRISPR-based technologies will aid in developing predictive understanding of how genome sequence leads to gene regulatory function.


Matrix metalloproteinase-3, vitamin D receptor gene polymorphisms, and occupational risk factors in lumbar disc degeneration.
Lumbar disc degeneration (LDD) is a process that begins early in life, contributing to the development of low back pain. LDD is a consequence of a variety of factors, and its etiology remains poorly understood. Objectives to investigate occupational and genetic risk factors inducing lumbar disc degeneration, and to evaluate the possible association of genetic polymorphisms of matrix metalloproteinase 3 (MMP-3) and vitamin D receptor (VDR) with the severity of LDD in an Egyptian population. A case control study involving 84 LDD and 60 controls was carried out. Five types of work related factors were investigated by questionnaire, complete neurological examination for all subjects and MRI for the cases. Polymerase chain reaction and restriction fragment length polymorphism methods were applied to detect polymorphisms in MMP-3 Promoter (-1,171 6A/5A) (rs 731236) and VDR-Apa (rs 35068180). We found that family history, back injury, smoking, high level of sitting, bending/twisting, physical workload, lifting, whole body vibration, mutant allele 5A of MMP-3 and mutant allele T of VDR were significantly associated with LDD (OR = 2.9, 3.1, 2.1, 11.1, 15.9, 11.7, 8.2, 12.6, 2.5 and 3.1 respectively, p < 0.05). Cases that carry allele 5A and/or allele T were associated with LDD severity. LDD is closely associated in occurrence and severity with occupational, environmental risk factors and susceptibility genes namely MMP-3, and VDR (ApaI). This study throws light on the importance of screening for early detection of susceptible individuals and disease prevention.


An alternative procedure for the creation of an ileal conduit in patients undergoing pelvic exenteration: dextrotransmesenteric location.
Ileal conduits have long been accepted as a standard method for urinary diversion, but conventional ileal conduits are not always suitable for patients whose ureters are for the greater part compromised by neoplasm or preoperative irradiation, resulting in a shortening of the ureters. Bowel migration into the large cavity, which develops after pelvic organ removal, appears to cause tension on the ureteroileal anastomotic site. Dextrotransmesenteric placement of an ileal conduit may provide easy access to the shortened ureters without exerting pressure on the anastomosis. We employed this procedure in seven patients undergoing pelvic exenteration with relatively minimal morbidity.


Left ventricular dysfunction in coronary artery disease: comparison between rest-redistribution thallium 201 and resting technetium 99m methoxyisobutyl isonitrile cardiac imaging.
We compared rest-redistribution thallium 201 and resting technetium 99m methoxyisobutyl isonitrile (MIBI) cardiac imaging in 29 men with angiographically proven coronary artery disease and regional ventricular dysfunction. Left ventricular ejection fraction at radionuclide angiography was 35% +/- 9%. Regional left ventricular wall motion was assessed on gated 99mTc MIBI images according to a 3-point scale (0 = normal, 1 = hypokinetic, 2 = akinetic or dyskinetic). 201Tl and 99mTc MIBI uptake values were analyzed quantitatively. A total of 435 myocardial segments were classified on the basis of wall motion analysis into three groups: group 1 (normal wall motion; n = 201), group 2 (hypokinetic; n = 132), and group 3 (akinetic or dyskinetic; n = 102). 201Tl and 99mTc MIBI uptake values were significantly higher in groups 1 and 2 compared with group 3 (p < 0.05) and in group 1 compared with group 2 (p < 0.05). When 201Tl and 99mTc MIBI uptake values were directly compared, no significant differences in group 1 and 2 were observed. In group 3, 99mTc MIBI uptake (67% +/- 14%) was significantly lower (p < 0.001) than initial (72% +/- 11%) and delayed 201Tl uptake (73% +/- 12%). Thus rest-redistribution 201Tl and resting 99mTc MIBI cardiac imaging reflect the severity of left ventricular dysfunction in coronary artery disease. However, in segments with severely impaired regional ventricular function, 201Tl uptake is significantly higher than 99mTc MIBI uptake.


A finite element model for ice ball evolution in a multi-probe cryosurgery.
The ice formation in a water body is examined for the computation of temperature field, phase change and a moving ice-water interface whose location is not known á priori. This is classically referred to as the Stefan problem [Rubinstein, L.I. (1971) The Stefan Problem (American Mathematical Society, Providence, Rhode Island 02904]. Based on the Duvaut [Duvaut, G. (1973) "Résolution d'un probléme Stefan" C.R. Acad Sci. Paris 276, 1461-1463] transformation, the governing equations for heat conduction are formulated within a variational principle that is readily amenable to a standard finite element solution without remeshing. Numerical simulation results pertaining to the freezing of tumour tissue in a multi-cryoprobe cryosurgery are presented. These results lend both quantitative and graphical support to the current empirical standards of "effective therapy" in view of refining clinical applications.


Seminal exosomes induce interleukin-6 and interleukin-8 secretion by human endometrial stromal cells.
Exosomes are extracellular microvesicles that participate in intercellular communication. Seminal plasma (SP) contains very large amounts of exosomes which are deposited in female genital tract after insemination. Although the response of vaginal cells to seminal exosomes (SE) is recently being elucidated, the interaction of uterine cells with SE is still unknown. Here, we aimed to evaluate the effect of SE on cytokine secretion by human endometrial stromal cells (eSC). Exosomes were isolated from the semen samples of healthy men with proven fertility and characterized using common exosome characterization methods. Human eSC were isolated from endometrial biopsies obtained from healthy premenopausal women. For exosome internalization analysis, SE were labeled with PKH67 green fluorescent dye and incubated with the cells. For investigating the effect of SE on cytokine secretion of eSC, we measured levels of interleukin (IL)-6, IL-8, IL-10, IL-1α, and leukemia inhibitory factor (LIF) in the culture supernatants of control and experimental groups by enzyme-linked immunosorbent assay (ELISA) after 24 h of incubation. Our results demonstrated that SE are internalized by eSC and subsequently induce them to produce IL-6 and IL-8, the cytokines which are involved in the immunology of embryo implantation. The findings of the present study suggest that SE contribute to the immunoregulatory functions of SP in the uterus and may participate in embryo implantation process. Therefore dysfunction of intracellular machineries of SE biogenesis and secretion, inadequate production, defective transportation to the uterus and impaired communication with endometrium may play a distinct role in pathophysiology of embryo implantation failure.


A global review on ambient Limestone-Precipitating Springs (LPS): Hydrogeological setting, ecology, and conservation.
Springs are biodiversity hotspots and unique habitats that are threatened, especially by water overdraft. Here we review knowledge on ambient-temperature (non-geothermal) freshwater springs that achieve sufficient oversaturation for CaCO3 -by physical CO2 degassing and activity of photoautotrophs- to deposit limestone, locally resulting in scenic carbonate structures: Limestone-Precipitating Springs (LPS). The most characteristic organisms in these springs are those that contribute to carbonate precipitation, e.g.: the mosses Palustriella and Eucladium, the crenophilous desmid Oocardium stratum, and cyanobacteria (e.g., Rivularia). These organisms appear to be sensitive to phosphorus pollution. Invertebrate diversity is modest, and highest in pools with an aquatic-terrestrial interface. Internationally, comprehensive legislation for spring protection is still relatively scarce. Where available, it covers all spring types. The situation in Europe is peculiar: the only widespread spring type included in the EU Habitat Directive is LPS, mainly because of landscape aesthetics. To support LPS inventorying and management to meet conservation-legislation requirements we developed a general conceptual model to predict where LPS are more likely to occur. The model is based on the pre-requisites for LPS: an aquifer lithology that enables build-up of high bicarbonate and Ca(2+) to sustain CaCO3 oversaturation after spring emergence, combined with intense groundwater percolation especially along structural discontinuities (e.g., fault zones, joints, schistosity), and a proper hydrogeological structure of the discharging area. We validated this model by means of the LPS information system for the Emilia-Romagna Region (northern Italy). The main threats to LPS are water diversion, nutrient enrichment, and lack of awareness by non-specialized persons and administrators. We discuss an emblematic case study to provide management suggestions. The present review is devoted to LPS but the output of intense ecological research in Central Europe during the past decades has clearly shown that effective conservation legislation should be urgently extended to comprise all types of spring habitats.


Exacerbation of liver cirrhosis in pregnancy: a complex emerging clinical situation.
Pregnancy in women with liver cirrhosis is an infrequent situation. In general the incidence of cirrhosis in pregnancy is very low and it is estimated approximately about 1 in 5,950 pregnancies. We report a descriptive management of pregnancy in a woman suffering from chronic viral hepatitis B with D and C coinfection exacerbated finally to severe cirrhosis of the liver. Caesarean section was performed giving birth to a viable male infant. Two months after the operation the patient underwent a liver transplantation with good outcome. Management of such severe cases requires a multidisciplinary medical approach. Data regarding cirrhosis caused by chronic viral agents in pregnancy are limited.


Phytochemical and biological studies on rare and endangered plants endemic to China. Part XV. Structurally diverse diterpenoids and sesquiterpenoids from the vulnerable conifer Pseudotsuga sinensis.
An extensive phytochemical investigation on the chemical constituents from the needles and twigs of the vulnerable conifer Pseudotsuga sinensis yielded 19 diterpenoids and 21 sesquiterpenoids with various carbocyclic skeletons. Among them, 13 (named pseudosinins A-M, resp.) were undescribed compounds. Their structures with absolute configurations were characterized by a combination of spectroscopic methods, calculated and experimental electronic circular dichroism (ECD) data, quantum chemical calculations of the chemical shifts, and single crystal X-ray diffraction analyses. In particular, an array of labdane-derived norditerpenoids with C19-, C18-, and C16-skeletons, and related drimane-type sesquitepenoids with C15- and C13-skeletons were found in the title plant. The possible biogenetic relationships of these degraded terpenoids were briefly discussed. Among the isolates, pseudosinin D, cis-communic acid, and 4β,15-dihydroxy-19-norabieta-8,11,13-trien-7-one showed moderate inhibitory activities against the enzyme ATP-citrate lyase (ACL), a potential drug target for the treatment of hyperlipidemia and hypercholesterolemia.


Pregnancy-Induced Hypertensive Disorders before and after a National Economic Collapse: A Population Based Cohort Study.
Data on the potential influence of macroeconomic recessions on maternal diseases during pregnancy are scarce. We aimed to assess potential change in prevalence of pregnancy-induced hypertensive disorders (preeclampsia and gestational hypertension) during the first years of the major national economic recession in Iceland, which started abruptly in October 2008. Women whose pregnancies resulted in live singleton births in Iceland in 2005-2012 constituted the study population (N = 35,211). Data on pregnancy-induced hypertensive disorders were obtained from the Icelandic Medical Birth Register and use of antihypertensive drugs during pregnancy, including β-blockers and calcium channel blockers, from the Icelandic Medicines Register. With the pre-collapse period as reference, we used logistic regression analysis to assess change in pregnancy-induced hypertensive disorders and use of antihypertensives during the first four years after the economic collapse, adjusting for demographic and pregnancy characteristics, taking aggregate economic indicators into account. Compared with the pre-collapse period, we observed an increased prevalence of gestational hypertension in the first year following the economic collapse (2.4% vs. 3.9%; adjusted odds ratio [aOR] 1.47; 95 percent confidence interval [95%CI] 1.13-1.91) but not in the subsequent years. The association disappeared completely when we adjusted for aggregate unemployment rate (aOR 1.04; 95% CI 0.74-1.47). Similarly, there was an increase in prescription fills of β-blockers in the first year following the collapse (1.9% vs.3.1%; aOR 1.43; 95% CI 1.07-1.90), which disappeared after adjusting for aggregate unemployment rate (aOR 1.05; 95% CI 0.72-1.54). No changes were observed for preeclampsia or use of calcium channel blockers between the pre- and post-collapse periods. Our data suggest a transient increased risk of gestational hypertension and use of β-blockers among pregnant women in Iceland in the first and most severe year of the national economic recession.


Resistance to Poly (ADP-Ribose) Polymerase Inhibitors (PARPi): Mechanisms and Potential to Reverse.
This review will focus on the most common mechanisms for poly (ADP-ribose) polymerase inhibitors' (PARPi) resistance and the main strategies for overcoming acquired or de novo PARPi resistance. Initial approvals for PARPi as part of treatment for advanced epithelial ovarian cancer (EOC) started in 2014 with patient with recurrent cancer characterized by BRCA mutations in the 3rd and 4th line and now have approvals for front-line maintenance in both the BRCA mutated and BRCAwt populations. As with all therapies, patients will eventually develop resistance to treatment. The most common mechanisms for PARPi resistance include reversion mutations, methylation events, and restoration of homologous recombination deficiency (HRD) through combinations and targeting replication stress. As more and more patients receive initial treatment (and potential retreatment with PARPi), we need to better understand the mechanisms in which tumors acquire PARPi resistance.


Contribution of vesicle trafficking dysregulation to the pathomechanism of mucopolysaccharidosis.
Although mucopolysaccharidoses (MPS) are monogenic diseases, caused by mutations in genes coding for enzymes involved in degradation of glycosaminoglycans (GAGs), recent studies suggested that changes in expressions of various genes might cause secondary and tertiary cellular dysfunctions modulating the course of these diseases. In this report, we demonstrate that vesicle trafficking regulation is affected in fibroblasts derived from patients suffering from 11 different types of MPS due to changes in levels of crucial proteins (estimated by automated Western-blotting) involved in this process, including caveolin, clathrin, huntingtin (Htt), APPL1, EEA1, GOPC, Rab5, and Rab7. Microscopic studies confirmed these results, while investigations of tissue samples derived from the MPS I mouse model indicated differences between various organs in this matter. Moreover, transcriptomic analyses provided a global picture for changes in expressions of genes related to vesicle trafficking in MPS cells. We conclude that vesicle trafficking is dysregulated in MPS cells and changes in this process might contribute to the molecular mechanisms of this disease. Most probably, primary GAG storage might cause a cellular stress response leading to dysregulation of expression of many genes which, in turn, results in changes in cellular processes like vesicle trafficking. This can significantly modulate the course of the disease due to enhancing accumulation of GAGs and altering crucial cellular processes. This hypothesis has been supported by normalization of levels of clathrin in MPS cells treated with either an active form of the deficient GAG-degrading enzyme or a compound (5,7-dihydroxy-3-(4-hydroxyphenyl)-4H-1-benzopyran-4-one) indirectly reducing the efficiency of GAG synthesis.


Integration of TGF-β-induced Smad signaling in the insulin-induced transcriptional response in endothelial cells.
Insulin signaling governs many processes including glucose homeostasis and metabolism, and is therapeutically used to treat hyperglycemia in diabetes. We demonstrated that insulin-induced Akt activation enhances the sensitivity to TGF-β by directing an increase in cell surface TGF-β receptors from a pool of intracellular TGF-β receptors. Consequently, increased autocrine TGF-β signaling in response to insulin participates in insulin-induced angiogenic responses of endothelial cells. With TGF-β signaling controlling many cell responses, including differentiation and extracellular matrix deposition, and pathologically promoting fibrosis and cancer cell dissemination, we addressed to which extent autocrine TGF-β signaling participates in insulin-induced gene responses of human endothelial cells. Transcriptome analyses of the insulin response, in the absence or presence of a TGF-β receptor kinase inhibitor, revealed substantial positive and negative contributions of autocrine TGF-β signaling in insulin-responsive gene responses. Furthermore, insulin-induced responses of many genes depended on or resulted from autocrine TGF-β signaling. Our analyses also highlight extensive contributions of autocrine TGF-β signaling to basal gene expression in the absence of insulin, and identified many novel TGF-β-responsive genes. This data resource may aid in the appreciation of the roles of autocrine TGF-β signaling in normal physiological responses to insulin, and implications of therapeutic insulin usage.


Targeting the resolution pathway of inflammation using Ac2-26 peptide-loaded PEGylated lipid nanoparticles for the remission of rheumatoid arthritis.
Rheumatoid arthritis (RA) is a common autoimmune disease characterized by joint inflammation and immune dysfunction. Although various therapeutic approaches have been utilized for the treatment of RA in clinical applications, the low responsiveness of RA patients and undesired systemic toxicity are still unresolved problems. Targeting the resolution pathway of inflammation with pro-resolving mediators would evoke the protective actions of patient for combating the inflammation. Ac2-26, a 25-amino acid peptide derived from Annexin A (a pro-resolving mediator), has shown good efficacy in the treatment of inflammatory disorders. However, the low bioavailability of Ac2-26 peptides hinders their efficacy in vivo. In this paper, we formed PEGylated lipid nanoparticles (LDNPs) by the co-assembly of l-ascorbyl palmitate (L-AP) and N-(carbonyl methoxypolyethylene glycol-2000)-1,2-distearoyl-sn‑glycero-3-phosphoethanolamine (DSPE-PEG2k) to encapsulate and deliver Ac2-26 peptides to the arthritic rats. They showed good stability and biocompatibility. After being intravenously administrated, Ac2-26 peptide-loaded PEGylated lipid nanoparticles (ADNPs) showed the prolonged in vivo circulation time and enhanced accumulation in inflamed sites. In vivo therapeutic evaluations revealed that ADNPs could attenuate synovial inflammation and improve joint pathology. Therefore, the pro-resolving therapeutic strategy using ADNPs is effective in RA treatment.


[Effect of nano-SiO2 exposure on spatial learning and memory and LTP of hippocampal dentate gyrus in rats].
To study the effect of nano-SiO2 on spatial learning and memory. Twenty-four male rats were randomly divided into 3 groups: control group (C group), low dose group (L group) and high dose group (H group). The rats were intragastrically administrated with nanometer particles at 25 and 100 mg/kg body weight every day for 4 weeks. After exposure, the ability of learning and memory of rats was tested by Morris water maze, and electrophysiological brain stereotactic method was used to test long-tear potentiation (LTP) in dentate gyrus (DG) of the rats. The increase rate of body weight in H group was reduced significantly compared with C group ( P < 0.05). In the space exploration experiment of Morris water maze test, the escape latency of H group was longer than that of C group (P < 0.05). The rats of H group spent less time in finding the target quadrant (P < 0.05) . The rate of LP induction of H group was significantly lower than that of C group (P < 0.05). After high fre quency stimulation (HFS), The changes of amplitude of population spike (PS) of L group and H group were lower than those of C group significantly (P < 0.05, P < 0.01). Nano-SiO₂may result in impairment of spatial learning and memory ability by reducing the rate of LTP induction and the increase of PS in hippocampus.


Characteristics of precipitation chemistry at Lushan Mountain, East China: 1992-2009.
Trends in precipitation pH and conductivity during 1992-2009, and in ionic compositions from January 2007 to June 2009, are reported from Lushan Mountain, one of the highest mountains in mid-east China. Annual mean pH was in the range of 4.35-5.01 and showed a statistically very significant (P < 0.01) decreasing trend with time. Annual mean conductivity showed a statistically significant (P < 0.05) increasing trend, although this was not the case for non-H conductivity. Increasing rainwater acidity was mainly caused by increasing amounts of acid substances entering the rain. The trends in precipitation pH and conductivity were directly associated with energy consumption. Over the period of study, Lushan Mountain received more rainfall in spring and summer. The pH values varied seasonally with winter minima. The winter multiyear seasonal mean pH was 4.35. The corresponding summer value was 4.88. SO (4) (2-) and NO (3) (-) were the main anions, and NH (4) (+) and Ca(2+) the main cations. The anion to cation ratio was 0.8-1.0, and that of [SO (4) (2-) ] to [NO (3) (-) ] was 2.4-3.0, much lower than that of the 1980s. However, sulfuric acid was still the main acid present. The ratio of [NH (4) (+) ] to [Ca(2+)] was about 1.0, suggesting that these two alkaline substances provided close acid neutralizing capacity. The ratio of [Cl(-)] to [Na(+)] was about 0.67, somewhat lower than that of natural precipitation. Ionic composition varied seasonally and was closely correlated to the amounts of rainfall and pollution. Trajectory analyses showed that the trajectories to Lushan Mountain could be classified in six clusters and trajectories originating from the South Sea and the areas surrounding Lushan Mountain had the greatest impacts on precipitation chemistry.


Localization of the regulatory subunit of cAMP-dependent protein kinase in Saccharomyces cerevisiae.
The subcellular distribution of the regulatory subunit of cAMP-dependent protein kinase in Saccharomyces cerevisiae cells was determined by subcellular fractionation and indirect immunofluorescence microscopy using the bcy1 mutant deficient in the regulatory subunit as control. The regulatory subunit of cAMP-dependent protein kinase showing cAMP-binding activity was identified as a single protein of 50 kDa by photoaffinity labeling and immunoblotting. The regulatory subunit was concentrated in a nuclear fraction in addition to a cytoplasmic fraction. By comparison of the regulatory subunit distribution with the DNA localization, the area detected by the indirect immunofluorescence was identified as the nucleus.


Histamine release in anaesthesia and surgery. premedication with H1- and H2-receptor antagonists: indications, benefits and possible problems.
Our clinical and experimental studies have so far demonstrated, that the drugs used in anaesthesia; such as hypnotics, sedatives, narcotics or muscle relaxants, release histamine. The intravenous short acting anaesthetic etomidate has not shown either in experimental studies or in clinical use for 10 years any anaphylactoid reaction. The benzodiazepines are another group of drugs which appear not to release high amounts of histamine. Accurate studies on volunteers as well as on patients on the application of H1- and H2-receptor antagonists have demonstrated an effective inhibition of the anaphylactoid reaction. We suggest that in case of a history of allergy H1- and H2-receptor antagonists should be administered as a prophylactic premedication.


Adrenal hydroxylations in genetically obese and hypertensive rats.
The separate steps in the formation of aldosterone from cholesterol were studied in a strain of spontaneously hypertensive rats in which phenotypic obesity is inherited as a recessive trait (Koletsky rats). The obese and hypertensive state had little or no effect on side-chain cleavage of cholesterol, formation of progesterone from pregnenolone or 21-hydroxylation. Mitochondrial 18-hydroxylation of endogenous and exogenous corticosterone, however, as well as 18- and 11 beta-hydroxylation of deoxycorticosterone, were increased in obese hypertensive rats, both when compared with non-obese hypertensive siblings and when compared with healthy Sprague-Dawley rats. 18-Hydroxylation of corticosterone was increased more than 18-hydroxylation of deoxycorticosterone. In non-obese hypertensive rats, the adrenal content of mitochondrial cytochrome P-450 was lower than that in obese hypertensive rats but higher than that in rats of the conventional Sprague-Dawley strain. The results are discussed with respect to possible heterogeneity of adrenal cytochrome P-450 and to possible explanations for the changes observed.