Non-invasive measurements of myocardial carbon metabolism using in vivo 13C NMR spectroscopy.
Despite their prime role in maintaining contractile performance, myocardial substrate uptake, substrate preference and metabolism are difficult to assess non-invasively. The objective of the present work was to extend the scope of cardiac 13C nuclear magnetic resonance (NMR) spectroscopy to the in vivo situation ('closed-chest model') and to quantitatively appraise myocardial metabolism in vivo. For this purpose, overnight-fasted Sprague-Dawley rats received intravenous infusions of non-radioactive 13C-labeled glucose, 3-hydroxybutyrate, and acetate as markers for glycolysis, metabolism of ketone bodies and direct incorporation into tricarboxylic acid (TCA) cycle, respectively. In vivo 13C NMR spectra (at 7 T) were acquired from the myocardium with a time resolution of 6 min. At the end of the infusion experiments, tissue extracts were prepared and further analyzed by high-resolution 13C NMR spectroscopy in order to corroborate the findings obtained in vivo. Accordingly, 3-hydroxybutyrate and acetate were rapidly extracted by the myocardium and supplied 42 +/- 6 and 53 +/- 9% of the acetyl-CoA for TCA cycle operation, whereas glucose, although also well extracted, did not contribute to myocardial oxidative metabolism. Myocardial TCA cycle turnover (V(TCA)) in vivo was estimated at 1.34 +/- 0.07 micromol/min/g wet weight, myocardial oxygen consumption (MVO2) at 2.95 +/- 0.16 micromol/min/g wet weight, exchange rate between alpha-ketoglutarate and glutamate (V(x)) at 1.22 +/- 0.08 micromol/min/g wet weight and rate of glutamine synthesis (V(gln)) at 0.14 +/- 0.02 micromol/min/g wet weight. The substantial synthesis of myocardial glutamine is in contrast to experiments with isolated and saline perfused hearts. In conclusion, it is demonstrated that 13C NMR spectroscopy of the heart in intact rats is feasible and provides new quantitative insight into myocardial substrate uptake, preference and metabolism in vivo.


["Body packing" in Austria. Clinical and diagnostic aspects of gastrointestinal smuggling of intoxicating drugs based on 2 cases].
The alimentary tract is frequently used as a hiding place for the transportation of drugs. We report on the cases of two smugglers with drug packages ("body packing") in the gastrointestinal tract. Radiological techniques provide the best method of diagnosis. The clinical problems and symptoms of acute drug intoxication are presented.


Gene structure evolution of the Na+-Ca2+ exchanger (NCX) family.
The Na+-Ca2+ exchanger (NCX) is an important regulator of cytosolic Ca2+ levels. Many of its structural features are highly conserved across a wide range of species. Invertebrates have a single NCX gene, whereas vertebrate species have multiple NCX genes as a result of at least two duplication events. To examine the molecular evolution of NCX genes and understand the role of duplicated genes in the evolution of the vertebrate NCX gene family, we carried out phylogenetic analyses of NCX genes and compared NCX gene structures from sequenced genomes and individual clones. A single NCX in invertebrates and the protochordate Ciona, and the presence of at least four NCX genes in the genomes of teleosts, an amphibian, and a reptile suggest that a four member gene family arose in a basal vertebrate. Extensive examination of mammalian and avian genomes and synteny analysis argue that NCX4 may be lost in these lineages. Duplicates for NCX1, NCX2, and NCX4 were found in all sequenced teleost genomes. The presence of seven genes encoding NCX homologs may provide teleosts with the functional specialization analogous to the alternate splicing strategy seen with the three NCX mammalian homologs. We have demonstrated that NCX4 is present in teleost, amphibian and reptilian species but has been secondarily and independently lost in mammals and birds. Comparative studies on conserved vertebrate homologs have provided a possible evolutionary route taken by gene duplicates subfunctionalization by minimizing homolog number.


Term extrauterine pregnancy in a Nigerian mother: a complication of uterine dehiscence.
Abdominal pregnancy is an uncommon but life-threatening form of ectopic pregnancy. It is associated with high maternal/fetal morbidity and mortality. We present a rare case of term abdominal pregnancy resulting from anterior uterine wall dehiscence, in a 36-year-old woman with three previous caesarean sections. The diagnosis was made at laparotomy for the fourth "caesarean section".


Total homocysteine and cognitive deterioration in people with type 2 diabetes.
The aim of this study was to investigate the relationship between total homocysteine levels in people with Type 2 diabetes and cognitive status. Fifty patients from our diabetes unit (30 females/20 males) with diabetes were enrolled. All patients had fasting blood samples taken for measurement of cardiovascular risk factors; total cholesterol and triglyceride concentrations and other lipid fractions (lipoprotein (a), low density lipoprotein (LDL-cholesterol), high density lipoprotein (HDL-cholesterol)), glucose, HbA(1c) and homocysteine. 24-h urine collection was used to measure creatinine clearance and microalbuminuria. Vitamin B-12 and folate were measured to assess vitamin status. All diabetic patients were assessed for late complications and a Mini-Mental State Examination (MMSE) was performed. The patients were 64.6 (49-78) years old with body mass index (BMI) of 29.6 +/- 6.3 kg/m(2), and duration of diabetes of 8.9 +/- 6.7 years. A univariant correlation analysis was performed among cardiovascular risk factors and vitamins with total MMSE score. Total homocysteine was inverse by correlated with MMSE score (r=-0.38; P<0.05) of the other measures of cardiovascular risk, microalbuminuria showed an inverse correlation with MMSE score (r=-0.51:P<0.01). Lipoproteins, glucose control and vitamin status were not correlated MMSE score. In the multiple regression model only microalbuminuria remained in the model, showing a decrease of one point in the MMSE result with each milligram of microalbuminuria, adjusted for confounding factors. Cognitive status in type 2 diabetic was correlated with homocysteine levels and microalbuminuria, this last endothelial damage marker remaining as an independent risk factor of cognitive deterioration.


Refractory response to entrectinib for ROS-1 rearranged NSCLC with concurrent de novo TP53 mutation showing good response to CNS lesion, but poor duration of response: A case report.
Entrectinib, a ROS-1 inhibitor, has been shown to be effective for patients with ROS-1 fused NSCLC, and has been established as the standard of care for this population. Entrectinib has been shown to achieve a better response to brain metastasis due to the characteristic of the drug having a weak interaction with P-glycoprotein and, even in prospective studies, the intracranial response is higher. Patients have been known to acquire resistance to molecularly targeted drugs such as EGF-TKIs or ALK-TKIs during targeted therapy. Similarly, the mechanisms of resistance to entrectinib have been reported, but information about the effects of TP53 mutation with entrectinib are still limited. Here, we experienced a case of a patient with ROS-1 fusion and concurrent TP53 mutation who was treated with entrectinib, resulting in a response to brain metastasis but rapid resistance to entrectinib. Our case demonstrates both the intracranial activity of entrectinib and the potential for resistance to entrectinib due to TP53 mutation.


Proteomic insights into floral biology.
The flower is the most important biological structure for ensuring angiosperms reproductive success. Not only does the flower contain critical reproductive organs, but the wide variation in morphology, color, and scent has evolved to entice specialized pollinators, and arguably mankind in many cases, to ensure the successful propagation of its species. Recent proteomic approaches have identified protein candidates related to these flower traits, which has shed light on a number of previously unknown mechanisms underlying these traits. This review article provides a comprehensive overview of the latest advances in proteomic research in floral biology according to the order of flower structure, from corolla to male and female reproductive organs. It summarizes mainstream proteomic methods for plant research and recent improvements on two dimensional gel electrophoresis and gel-free workflows for both peptide level and protein level analysis. The recent advances in sequencing technologies provide a new paradigm for the ever-increasing genome and transcriptome information on many organisms. It is now possible to integrate genomic and transcriptomic data with proteomic results for large-scale protein characterization, so that a global understanding of the complex molecular networks in flower biology can be readily achieved. This article is part of a Special Issue entitled: Plant Proteomics--a bridge between fundamental processes and crop production, edited by Dr. Hans-Peter Mock.


Management of complications of periarticular fractures of the distal interphalangeal, proximal interphalangeal, metacarpophalangeal, and carpometacarpal joints.
Fractures involving the bones of the hand are among the most common injuries in the United States. A significant portion of these fractures are periarticular. Although the great majority of these fractures are treated successfully by nonoperative means, complications arise. We present a comprehensive review of prevention and management of complications of periarticular fractures of the distal interphalangeal, proximal interphalangeal, metacarpophalangeal, and carpometacarpal joints.


Neurocognitive impairment and gray matter volume reduction in HIV-infected patients.
Although neuropsychological studies of human immunodeficiency virus (HIV)-infected patients have demonstrated heterogeneity in neurocognitive impairment and neuroimaging studies have reported diverse brain regions affected by HIV, it remains unclear whether individual differences in neurocognitive impairment are underpinned by their neural bases. Here, we investigated spatial distribution patterns of correlation between neurocognitive function and regional gray matter (GM) volume across patients with HIV. Thirty-one combination antiretroviral therapy-treated HIV-infected Japanese male patients and 33 age- and sex-matched healthy controls were included in the analysis after strict exclusion criteria, especially for substance use. Fifteen neurocognitive tests were used, and volumetric magnetic resonance imaging was performed. We used voxel-based morphometry to compare GM volume between groups and identify regional GM volumes that correlated with neurocognitive tests across patients. Using the Frascati criteria, 10 patients were diagnosed with asymptomatic neurocognitive impairment, while the others were not diagnosed with HIV-associated neurocognitive disorders. Patients showed a significantly lower performance in five neurocognitive tests as well as significantly reduced GM volume relative to controls, with volume-reduced regions spread diffusely across the whole brain. Different aspects of neurocognitive impairment (i.e., figural copy, finger tapping, and Pegboard) were associated with different GM regions. Our findings suggest a biological background constituting heterogeneity of neurocognitive impairment in HIV infection and support the clinical importance of considering individual differences for tailor-made medicine for people living with HIV.


Pharmacokinetics and Bioequivalence Evaluation of a New Oxycodone Tamper-Resistant Tablet Administered with an Opioid Antagonist in Patients with Chronic Pain.
Oxycodone tamper resistant (OTR) is a new extended-release abuse-deterrent formulation providing improvements in the tamper resistant characteristics. This study aimed to investigate the pharmacokinetic properties of the new OTR tablets and evaluate the bioequivalence of oxycodone from OTR and the original extended release (ER) formulation tablets administered with an opioid antagonist in patients with chronic pain. In this open-label, randomized, cross-over study, the enrolled patients were randomised to receive a single dose of 40 mg OTR or 40 mg OXYCONTIN® (OXY) tablet administered with naltrexone blockade under fasting conditions. Serial blood samples for pharmacokinetic analysis were collected. Plasma oxycodone was quantified by a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method. Tolerability was evaluated by monitoring adverse events, physical examinations, 12-lead ECG and laboratory tests. A total of 38 patients were enrolled and 33 subjects completed the study. After a single dose of 40 mg tablets, pharmacokinetic results of the new OTR tablet were found to be similar to those of original extended-release oxycodone tablet. OTR 40 mg was bioequivalent to OXY 40 mg and was well tolerated in patients with chronic pain. The new OTR formulation could provide a new choice in the treatment of chronic pain and reduce the potential for oxycodone abuse. Chictr.org identifier: ChiCTR1800017253.


Fitness Mediates Activity and Sedentary Patterns Associations with Adiposity in Youth.
We aimed to examine the cross-sectional associations of patterns of sedentary time (ST) and moderate-to-vigorous physical activity (MVPA) with total fat mass index (FMI) and abdominal FMI (FMIabd) in children, and additionally, to analyze if cardiorespiratory fitness (CRF) mediated these associations. Fat mass index and FMIabd were assessed with dual-energy X-ray absorptiometry in 333 participants (172 girls) age 9 to 11 yr. MVPA, ST, breaks in ST per sedentary hour (BST/ST) and the number of daily sedentary bouts with various lengths (1-4, 5-9, 10-14, and ≥15 min) were assessed with accelerometry and CRF using a maximal cycle test. Hayes' PROCESS macro for SPSS was used for mediation analysis. The number of shorter sedentary bouts (1-4 min) was inversely associated with FMI (β = -0.108), whereas longer sedentary bouts (5-9 min, β = 0.169; 10-14 min, β = 0.193; ≥15 min, β = 0.377) had a positive association, independent of MVPA (P < 0.05). A similar trend was found for FMIabd. Moderate-to-vigorous physical activity was negatively related to FMI (β = -0.029) and to FMIabd (β = -0.003). There were no main associations for ST and BST/ST (P > 0.05). Cardiorespiratory fitness was inversely related to total and abdominal adiposity, and a large portion (40.9-65.7%) of the associations of MVPA and sedentary bouts with both FMI and FMIabd were mediated by CRF. Time in MVPA and accumulating ST in shorter sedentary bouts is inversely related to adiposity. However, these associations may be mediated by CRF. Emphasis should be given to programs that target increases in MVPA, which may influence CRF while reducing sedentary recreational behaviors.


Long-term visual, anterior and posterior corneal changes after crosslinking for progressive keratoconus.
To evaluate the long-term outcomes of corneal cross-linking (CXL) in patients with progressive keratoconus. In this retrospective non-comparative study, forty-five eyes of 31 patients with progressive keratoconus were treated with 30 min "epi-off" corneal cross-linking. The visual, refractive, topographic and tomographic outcomes were evaluated preoperatively and at least 10 years after cross-linking. Ten years post-corneal cross-linking, the mean anterior maximum keratometry decreased (-2.10 ± 3.25 D, P = 0.0001). Conversely, the posterior maximum keratometry increased (6.38 ± 9.79 D, P = 0.065). Both uncorrected and corrected distance visual acuity improved (LogMAR -0.08 ± 0.30) and (LogMAR -0.05 ± 0.21), respectively (P > 0.05, both). A statistically significant hyperopic shift was observed postoperatively (0.70 ± 1.31 D, P = 0.0009). The anterior topographic cylinder values revealed no change (-0.17 ± 1.31 D, P = 0.3), whereas the mean posterior cylinder values decreased (absolute value increased) significantly compared to baseline from -1.31 ± 0.97 D to -1.82 ± 1.78 D, (P < 0.05). The minimum corneal thickness values decreased significantly (-35.11 ± 48.63 µm, P = 0.0001). Four eyes (8.8%) showed more than 1 D increase in the anterior maximum keratometry. This protocol and duration of Epi-off corneal cross-linking was found to be effective in halting keratoconus progression over the follow up period (10 years). Anterior corneal flattening and a hyperopic shift were observed. A statistically significant increase in the posterior corneal cylinder was observed. Although, not reaching statistical significance, the logMAR uncorrected and corrected visual acuity were improved.


New generation hydroxypropyl methylcellulose (HPMC-Ophtal H) for intraocular surgery: a confocal laser scanning microscopy study.
To determine the abundance and variety of particulate debris in new generation hydroxypropyl methylcellulose (HPMC) samples. Three samples of sterile HPMC-Ophtal H were examined using a confocal laser scanning microscope, scanning electron microscope and mass spectrometry. Confocal laser scanning microscopy of the samples showed that particulate matter in HPMC was less than 30 microns in size and at a density of less than 1000 particles per millilitre. No discernible details were seen by the scanning electron microscope, and mass spectrometry did not identify and bands. Improved methods of preparation including filtration may be responsible for decreased particulate debris in new generation HPMC.


Separation of Fc-binding serum proteins by preparative flat-bed isotachophoresis.
Preparative flat-bed isotachophoresis with discrete spacers was applied as a single-step procedure to separate 2 Fc-dependent activities of normal chicken serum, i.e., (1) the ability to raise the titre of haemagglutinating allo-antisera which is due to a high molecular weight beta-globulin (HEF), (2) the ability to activate guinea pig complement components in mixed complement reaction. The results demonstrate that the 2 activities can be clearly separated, and HEF must therefore be different from the first complement factor in the chicken. Under the chosen conditions the molecule active in the mixed complement reaction is not stacked in contrast to other serum protein including HEF. The same technique with human serum shows that human Clq behaves in the same was as the chicken complement factor. This means that by selective unstacking, flat-bed isotachophoresis can be used as an efficient single-step purification method for human and chicken Clq.


Circulating thrombopoietin and interleukin-6 in newly diagnosed autoimmune versus aplastic thrombocytopenia.
Circulating thrombopoietin and interleukin-6 concentrations were investigated in two different settings of thrombocytopenia. Twenty patients with autoimmune thrombocytopenic purpura (ATP), 12 patients with aplastic anemia (AA) and 15 healthy subjects were studied. Thrombopoietin was significantly increased in AA and deficient in ATP. Interleukin-6 was significantly increased in ATP, compared to both other groups.


Persistence of complexed acidic phospholipids in rapidly mineralizing tissues is due to affinity for mineral and resistance to hydrolytic attack: in vitro data.
Acidic phospholipids, complexed with calcium and inorganic phosphate, are components of extracellular matrix vesicles. Both the complexed acidic phospholipids and matrix vesicles have previously been shown to serve as hydroxyapatite (HA) nucleators in solution and when implanted in a muscle pouch. The present study supplies evidence that complexed acidic phospholipids can persist in mineralizing tissues both because of their affinity for HA and because of their resistance to hydrolysis by phospholipase A2. Calcium-phosphatidylserine-phosphate complex (CPLX-PS) synthesized with 14C-labeled phosphatidylserine (PS) was used to measure CPLX-PS affinity for HA using a Langmuir adsorption isotherm model. The affinity was shown to be higher and more specific than that of PS itself (K = 8.66 ml/mumol; N, the number of binding sites = 20.4 mumol/m2 as compared with previously reported values for PS of K = 3.33 ml/mumol, and N = 4.87 mumol/m2). Incorporated into synthetic liposomes and incubated in a calcium phosphate solution in which mineralization is induced by an ionophore, CPLX-PS showed behavior distinct from free PS. As previously reported, PS in these liposomes totally blocked HA formation. On the other hand, CPLX-PS in similar concentrations had a varied response, having no effect, slightly inhibiting, or actually promoting HA formation. CPLX-PS was also shown to be a poorer substrate for phospholipase A2 than PS, with Km = 4.63 mM for CPLX-PS and Km = 0.27 mM for PS; and Vmax = 0.029 ml/minute for CPLX-PS and Vmax = 0.066 ml/minute for PS. These data explain how complexed acidic phospholipids may persist in the growth plate and facilitate initial mineral deposition.


Iodine supplementation during pregnancy: a public health challenge.
Iodine deficiency remains the most frequent cause worldwide, after starvation, of preventable mental retardation in children. It causes maternal hypothyroxinemia, which affects pregnant women even in apparently iodine-sufficient areas, and often goes unnoticed because L-thyroxine (T4) levels remain within the normal range, and thyroid-stimulating hormone (TSH) is not increased. Even a mild hypothyroxinemia during pregnancy increases the risk of neurodevelopmental abnormalities, and experimental data clearly demonstrate that it damages the cortical cytoarchitecture of the fetal brain. The American Thyroid Association (ATA) recommends a supplement of 150 microg iodine/day during pregnancy and lactation, in addition to the use of iodized salt. We discuss the importance of iodine supplementation to ensure adequate T4 levels in all women who are considering conception and throughout pregnancy and lactation.


Use of a donor heart that had undergone previous cardiac surgery for ASD closure.
We report the use of a heart from a 19-year-old female donor who had undergone open heart surgery for closure of an atrial septal defect at the age of 12. The explantation procedure was modified to prevent hemodynamic instability. The recipient, a 59-year-old male with increased pulmonary vascular resistance had an uneventful recovery after transplantation.


Treatment of gastrointestinal autonomic neuropathy.
The symptoms caused by gastrointestinal autonomic neuropathy in diabetes mellitus is important to highlight since it affects a large proportion of people with diabetes, regardless of whether this is type 1 or type 2. Gastroparesis and general signs of bowel dysfunction, such as constipation, diarrhoea and abdominal pain are most often encountered and involve both pharmacological and non-pharmacological treatment options. This mini-review summarises a presentation given at the 'Diagnosis and treatment of autonomic diabetic neuropathy in the gut' symposium at the 2015 annual meeting of the EASD. It is accompanied by another mini-review on a topic from this symposium (by Azpiroz and Malagelada, DOI: 10.1007/s00125-015-3831-1 ) and a commentary by the Session Chair, Péter Kempler (DOI: 10.1007/s00125-015-3826-y ).


In vivo anaphylaxis in the rat: effects of phosphodiesterase inhibitors.
On the basis of their inhibitory capacities on the phosphodiesterase enzyme system, we studied the anti-anaphylactic effect of milrinone and sulmazole in comparison with theophylline. For this purpose anaphylactic shock was induced in actively sensitised, spontaneously breathing rats. Milrinone, sulmazole and theophylline reduced anaphylactic bronchoconstriction without affecting the antigen induced fall in blood pressure. Surprisingly, sulmazole reduced mortality significantly.


The cholinergic neurotoxin ethylcholine mustard aziridinium (AF64A) induces an increase in MAO-B activity in the rat brain.
Recently it was reported that there is an increase in monoamine oxidase B (MAO-B) activity in post-mortem brains of patients with Alzheimer's disease. It was postulated that this increase in MAO-B activity was due to gliosis associated with neuronal degeneration. The aim of the present investigation was to evaluate the effect on MAO of neuronal degeneration primarily affecting the cholinergic system. The specific cholinergic toxin AF64A (3 and 4.5 nmol) was injected bilaterally into the cerebral ventricles of rats. We then estimated MAO-A, MAO-B, dopamine (DA) uptake rates and choline acetyltransferase (ChAT) activities in hippocampus, striatum and cortex, 1, 2.5 and 4.5 weeks after the injection. Marked long-lasting reduction in ChAT activities appeared only in hippocampus, consistent with previous reports. The MAO-A activity was unchanged as were DA uptake rates. Neither was there any change in MAO-B activity found 1 week after the injection. However, a significant increase in MAO-B activity appeared after 2.5 weeks and persisted after 4.5 weeks in all 3 brain regions investigated. This result is likely to reflect progressive gliosis after cholinergic neuronal degeneration. Previous results have shown an increased MAO-B activity with age and a further accelerated increase in Alzheimer's disease. Experimentally, hemitransection and injection of kainic acid have been shown to cause a similar increase. The present results show that changes in MAO-B activity also reflect degenerative processes in brain mainly affecting the cholinergic system.


Study of different doses of zinc oxide nanoparticles by intraperitoneal and gavage methods on testicular tissue in Wistar rats: An experimental study.
Zinc oxide nanoparticles (ZnO-NPS) are widely used inhuman life; however, they do have side effects on human health. This study aimed to evaluatethedifferent doses of ZnO-NPS on testicular tissue. 35 male Wistar rats(10-12 wk, 220 ± 20gr) were divided into 7 groups of 5, including the control group (gavaged distilled water daily), sham group (received intraperitoneal doses of distilled water twice a week). The group receivedintraperitoneal ZnO-NPS (25, 50, and 100 mg/kg body weight, twice a week), and gavage (150 and 200 mg/kg body weight daily). All stages of the test were performed in 4 wk then serum testosterone and tissue malondialdehyde, and ferric reducing antioxidant power levels were measured, also testes histopathological evaluation was performed. Our results showed that a reduced cell population of spermatozoa was observed in the group that received 25 mg/kg ZnO-NPS, while a reduced cell population of spermatozoa, edema, hyperemia, and vacuolar degeneration were observed in the group that received 50 and 100 mg/kg ZnO-NPS. The maximum amount of lesions were observed in the dose of 200 mg/kg. The highest amount of ferric reducing antioxidant power and testosterone levels were observed in the control group. Also, a 100 mg/kg intraperitoneal dose of ZnO-NPS and 150 mg/kg oral dose of ZnO-NPS were suitable doses to create a model of male genital lesions. Nanoparticles are harmful factors for the reproductive system and consequently affect infertility, which requires the toxicity of the concentration of these nanoparticles to be evaluated and controlled.


Low-penetrance susceptibility to breast cancer due to CHEK2(*)1100delC in noncarriers of BRCA1 or BRCA2 mutations.
Mutations in BRCA1 and BRCA2 confer a high risk of breast and ovarian cancer, but account for only a small fraction of breast cancer susceptibility. To find additional genes conferring susceptibility to breast cancer, we analyzed CHEK2 (also known as CHK2), which encodes a cell-cycle checkpoint kinase that is implicated in DNA repair processes involving BRCA1 and p53 (refs 3,4,5). We show that CHEK2(*)1100delC, a truncating variant that abrogates the kinase activity, has a frequency of 1.1% in healthy individuals. However, this variant is present in 5.1% of individuals with breast cancer from 718 families that do not carry mutations in BRCA1 or BRCA2 (P = 0.00000003), including 13.5% of individuals from families with male breast cancer (P = 0.00015). We estimate that the CHEK2(*)1100delC variant results in an approximately twofold increase of breast cancer risk in women and a tenfold increase of risk in men. By contrast, the variant confers no increased cancer risk in carriers of BRCA1 or BRCA2 mutations. This suggests that the biological mechanisms underlying the elevated risk of breast cancer in CHEK2 mutation carriers are already subverted in carriers of BRCA1 or BRCA2 mutations, which is consistent with participation of the encoded proteins in the same pathway.


[Study of the reaction between steroids and serum albumin by the spin probe method].
The data obtained from the study of interaction of two steroids with serum albumin shows that D-ring of steroid molecule has more hidrophobic environment then A-ring in the complexes of steroid with protein. The plane of steroid is perpendicular to the long axis of protein molecule.


Halogenated compounds as inducers of lipid peroxidation in tissue slices.
Twenty-seven halogenated compounds were screened as potential inducers of lipid peroxidation in rat liver, kidney, spleen, and testes slices. In addition to the known lipid peroxidation inducers--carbon tetrachloride and bromotrichloromethane--the novel compounds carbon tetrabromide, p-bromobenzyl bromide, and benzyl bromide increased lipid peroxidation in each of the tissues studied. Lipid peroxidation was measured by release of thiobarbituric acid-reactive substances (TBARS) from the tissue slices. The amount of TBARS released from liver slices incubated with bromotrichloromethane, carbon tetrabromide, dichloromethane, bromobenzene, chloroform, bromoform, benzyl chloride, bromochloromethane, and carbon tetrabromide correlated with the lethality of these compounds as evaluated by their oral LD50 in rats. The lethality of a number of the compounds tested did not correlate with their capacity to induce lipid peroxidation.


The Presence of Cu Facilitates Adsorption of Tetracycline (TC) onto Water Hyacinth Roots.
Batch experiments were conducted to investigate the adsorption characteristics of tetracycline (TC), and the interactive effects of copper (Cu) on the adsorption of TC onto water hyacinth roots. TC removal efficiency by water hyacinth roots was ranging from 58.9% to 84.6%, for virgin TC, 1:1 TC-Cu and 1:2 TC-Cu. The Freundlich isotherm model and the pseudo-second-order kinetic model fitted the adsorption data well. Thermodynamics parameters ΔG⁰ for TC were more negative in the TC plus Cu than the TC-only treatments, indicating the spontaneity of TC adsorption increased with increasing of Cu concentrations. An elevated temperature was associated with increasing adsorption of TC by water hyacinth roots. The additions of Cu(II) significantly increased TC adsorption onto water hyacinth roots within the pH range 4 to 6, because copper formed a strong metal bridge between root surface and TC molecule, facilitating the adsorption of TC by roots. However, Cu(II) hindered TC adsorption onto water hyacinth roots on the whole at pH range from 6⁻10, since the stronger electrostatic repulsion and formation of CuOH⁺ and Cu(OH)₂. Therefore, the interaction between Cu(II) and TC under different environmental conditions should be taken into account to understand the environmental behavior, fate, and ecotoxicity of TC.


Impact of ambient and supplemental ultraviolet-B stress on kidney bean plants: an insight into oxidative stress management.
In the present study, the response of kidney bean (Phaseolus vulgaris L. cv. Pusa Komal) plants was evaluated under three different levels of ultraviolet-B (UV-B), i.e., excluded UV-B (eUV-B), ambient UV-B (aUV-B; 5.8 kJ m(-2) day(-1)), and supplemental UV-B (sUV-B; 280-315 nm; ambient + 7.2 kJ m(-2) day(-1)), under near-natural conditions. eUV-B treatment clearly demonstrated that both aUV-B and sUV-B are capable of causing significant changes in the plant's growth, metabolism, economic yield, genome template stability, total protein, and antioxidative enzyme profiles. The experimental findings showed maximum plant height at eUV-B, but biomass accumulation was minimum. Significant reductions in quantum yield (Fv/Fm) were observed under both aUV-B and sUV-B, as compared to eUV-B. UV-B-absorbing flavonoids increased under higher UV-B exposures with consequent increments in phenylalanine ammonia lyase (PAL) activities. The final yield was significantly higher in plants grown under eUV-B, compared to those under aUV-B and sUV-B. Total protein profile through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and analysis of isoenzymes, like superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), and glutathione reductase (GR), through native PAGE revealed major changes in the leaf proteome under aUV-B and sUV-B, depicting induction of some major stress-related proteins. The random amplified polymorphic DNA (RAPD) profile of genomic DNA also indicated a significant reduction of genome template stability under UV-B exposure. Thus, it can be inferred that more energy is diverted for inducing protection mechanisms rather than utilizing it for growth under high UV-B level.


Identification of Small Molecules that Modulate Mutant p53 Condensation.
Structural mutants of p53 induce global p53 protein destabilization and misfolding, followed by p53 protein aggregation. First evidence indicates that p53 can be part of protein condensates and that p53 aggregation potentially transitions through a condensate-like state. We show condensate-like states of fluorescently labeled structural mutant p53 in the nucleus of living cancer cells. We furthermore identified small molecule compounds that interact with the p53 protein and lead to dissolution of p53 structural mutant condensates. The same compounds lead to condensation of a fluorescently tagged p53 DNA-binding mutant, indicating that the identified compounds differentially alter p53 condensation behavior depending on the type of p53 mutation. In contrast to p53 aggregation inhibitors, these compounds are active on p53 condensates and do not lead to mutant p53 reactivation. Taken together our study provides evidence for structural mutant p53 condensation in living cells and tools to modulate this process.


Efficacy of Anterior Quadratus Lumborum Block and Pain After Total Hip Arthroplasty: A Randomized Controlled Trial.
Anterior quadratus lumborum block (AQLB) is one of the compartment blocks and has recently attracted attention as a new method of analgesia for postoperative hip surgery analgesia. This study aimed to compare the analgesic efficacy of AQLB in patients undergoing primary total hip arthroplasty (THA). There were 120 patients undergoing primary THA under general anesthesia randomly allocated to receive a femoral nerve block (FNB) or an AQLB. The primary outcome was total morphine consumption over the initial 24-hour postoperative period. Secondary outcomes included the pain score evaluation while at rest and during active and passive motion over the 2 days following surgery and the manual muscle testing of the quadriceps femoris. The numerical rating scale (NRS) score was used for evaluating the postoperative pain score. There were no significant differences between the 2 groups concerning morphine consumption within 24 hours after surgery (P = .72). The NRS score at rest and passive motion were similar at all-time points (P > .05). However, there was a statistically significant difference in pain reported during the active motion for the FNB group compared to the AQLB (P = .04). No significant differences were found between the 2 groups concerning muscle weakness incidence. Both AQLB and FNB demonstrated adequate efficacy for postoperative analgesia at rest in THA. However, based on our study, whether AQLB is inferior or noninferior to FNB as an analgesic method for THA was inconclusive.


Pressure release technique versus placebo applied to cervical and masticatory muscles in patients with chronic painful myofascial temporomandibular disorder: A randomised clinical trial.
The therapeutic approach to myofascial TMD should focus on pain relief and rehabilitation of function. This study investigated whether pressure release technique (PRT) is effective for reducing pain in people with chronic myofascial temporomandibular disorders (TMD). A single-blinded randomised parallel-group trial, with 3 months follow-up was conducted. A total of 72 patients were randomly allocated to receive PRT or sham PRT. Primary outcome was pain assessed with a visual analogue scale (VAS). Secondary outcomes included pressure pain thresholds (PPTs), range of opening of the mouth (ROM), Neck Disability Index (NDI), Pain Catastrophizing Scale (PCS), Tampa Scale for Kinesiophobia (TSK-11), State-Trait Anxiety Index (STAI) and State-Trait Depression Index (ST-DEP). All parameters were assessed at baseline, at the end of the treatment and at 3 months follow-up. Statistical analysis was performed by ANOVA. There were significant main effects of time, group and interaction between time and group (F ≥ 21.92; p < .001) on VAS pain. Post hoc tests showed a significant reduction in VAS pain scores in the PRT group (≥31.9%; p < .001). Effect sizes were moderate in the PRT group at all follow-up periods (≥1.25 Cohen's d). Also, there were significant effects of time in secondary outcomes (F ≥ 9.65; p < .001), and there were also interactions between time and group (F ≥ 3.82; p < .002) with better effects in the PRT group. The inclusion of PRT to conventional management with occlusal splints and self-care management appears to be effective to improve self-reported levels of pain in patients with chronic myofascial TMD pain. Retrospectively registered (ClinicalTrials.gov: NCT03619889).


Effectiveness of Bariatric Surgery in Patients with the Metabolically Healthy Obese Phenotype.
To evaluate the effectiveness and safety of bariatric surgery in metabolically healthy obese (MHO) patients. In this retrospective, observational study, we reviewed the medical records of patients who underwent bariatric surgery at a tertiary care hospital between January 2007 and March 2015. Patients who underwent revisional surgery and patients with type 1 diabetes were excluded from the analysis. MHO patients were defined as those without a previous diagnosis of diabetes or atherogenic dyslipidemia and absence of hypoglycemic treatment or treatment with fibrates. A total of 188 patients were included (mean age 48.97 ± 10.32 years, 68.6% of women). Sleeve gastrectomy was performed in 121 patients (64%) and a gastric bypass in 67 patients (36%). Prior to surgery, 36 patients (19%) were MHO. In the second- and third-year post-surgery, MHO patients presented a higher percentage of total weight loss (%TWL) (35.16% vs. 30.34%; p = 0.02 and 33.97% vs. 27.78%; p = 0.013 respectively). Multiple regression analysis showed that MHO was associated with a higher weight loss irrespective of age, sex, baseline BMI, and type of surgery. We did not detect any differences in acute complications between patients with and without MHO after bariatric surgery. Bariatric surgery in MHO patients in our study was associated with higher weight loss than that in MUHO patients. There were no differences between the two groups in respect to acute complications following surgery.


The Design of an Adaptive Clinical Trial to Evaluate the Efficacy of Extra-Corporeal Membrane Oxygenation for Out-of-Hospital Cardiac Arrest.
Extracorporeal cardiopulmonary resuscitation (ECPR) is a promising therapy for out-of-hospital cardiac arrest (OHCA) that is refractory to standard therapy, but no multicenter randomized clinical trials have been conducted to establish its efficacy. We report the design and operating characteristics of a proposed randomized Bayesian adaptive "enrichment" clinical trial designed to determine whether ECPR is effective for refractory OHCA and, if effective, to define the interval after arrest during which patients derive benefit. Through iterative trial simulation and trial design modification, we developed a Bayesian adaptive trial of ECPR for adults who experience non-traumatic out-of-hospital cardiac arrest. Our proposed trial design addresses the threats to trial success identified during the design process, which were (1) the uncertainty surrounding the cardiac arrest (CA)-to-ECPR interval within which clinical benefit might be preserved (2) the difference in prognosis between patients with an initial rhythm that is non-shockable vs. shockable. Trial subjects will be randomized 1:1 to receive either standard care or expedited transport to a hospital for potential ECPR. The CA-to-ECPR interval will be estimated in real time based on the sum of the estimated paramedic response time (911 call to scene arrival), paramedic scene time, and transport time to hospital. A Bayesian decreasing step function will be used to estimate the efficacy of the treatment with an outcome of the 90-day utility-weighted Modified Rankin Scale (uwmRS) for each rhythm subgroup and estimated CA-to-ECPR interval at pre-specified interims. The trial will adaptively lengthen the estimated CA-to-ECPR eligibility window if the treatment appears effective at the upper limit of initial eligibility window. If ECPR appears ineffective at longer estimated CA-to-ECPR intervals, the upper limit of the window for enrollment eligibility will be shortened. The analysis will be stratified by rhythm subgroup. With a maximum total sample size of 400, and a cap on the maximum sample size of 300 for the non-shockable rhythm subgroup, the trial design has power ranging from 91-100% to detect a benefit from ECPR for non-shockable rhythms under the various efficacy scenarios simulated and power ranging from 69-98% for shockable rhythms under the same scenarios. The trial design also has a high probability of correctly identifying the maximum CA-to-ECPR interval within which ECPR produces a clinically significant benefit of 0.2 on the uwMRS. If ECPR is equivalent to standard CA care, the type I error is 2.5% with a 99% probability of stopping enrollment early for futility in the non-shockable subgroup and a 97% probability of stopping enrollment early for futility in the shockable subgroup. This proposed adaptive trial design helps to ensure the population of patients who are most likely to benefit from treatment-as defined both by rhythm subgroup and estimated CA-to-ECPR interval-is enrolled. The design promotes early termination of the trial if continuation is likely to be futile.


Hydroxyl radical oxidation of cyclic methylsiloxanes D4 ∼ D6 in aqueous phase.
Cyclic methylsiloxanes (CMS) were listed as candidates of substances of very high concerns in 2018 by the REACH. These compounds can enter environmental waters, and potentially cause harmful effects to aquatic organisms and human beings. Until now, reaction mechanisms of these pollutants with hydroxyl radicals (HO) in aqueous phase were unknown. In this study, reaction mechanisms of three typical CMS (D4 ∼ D6) with HO in aqueous phase were investigated by employing both UV/H2O2 experiments and density functional theoretical calculations. Bimolecular reaction rate constants (kHO·) of D4 ∼ D6 with HO were determined as kHO·(D4) = 8kHO·(D5) = 12kHO·(D6) = 6.6 × 108 L mol-1 s-1. Half-lives of HO oxiding D4 ∼ D6 ranged from 12 to 140 days at [HO] = 10-15 mol L-1 in sunlit surface water, and were comparable to (D4, D5) or much shorter (D6) than hydrolytic half-lives. The reactivity to HO decreased with the increasing size of siloxane ring in aqueous phase, in an order totally opposite to that in gaseous phase. Calculation results indicated that HO oxidation of the three CMS proceeded spontaneously through an exothermic H-abstraction process at the first step. Water molecules participated into H-abstraction of CMS and caused energy barrier of D5 higher than that of D4. Thus, H-bonds formed by water molecules were responsible for the reverse reactivity of CMS in aqueous phase. This work provided basic evidences suggesting environmental persistence of CMS in aqueous phase completely different from that in gaseous phase.


Characterization of Vibrio harveyi strains recovered from diseased farmed Senegalese sole (Solea senegalensis).
To characterize 16 Vibrio harveyi strains isolated from different epizootic outbreaks affecting farmed Senegalese sole. The Vibrio harveyi strains tested have broad phenotypic diversity based on their biochemical and exoenzymatic patterns, outer membrane proteins (OMP), extracellular product (ECP) patterns and presence of prophages. Lethal dose 50 (LD(50)) of the strains and in vitro antagonism tests with two probiotic strains were also determined. The OMP analysis revealed three different patterns (A, M and V). The electrophoretic analysis of the ECP showed two different groups. All strains considered virulent based on their LD(50) exhibited the same protein pattern in their ECP (pattern I), while all nonvirulent strains showed a different profile (pattern II). About 32% of the tested strains were positive for prophages, although a clear relationship between virulence and the presence of prophages has not been established. The results obtained have shown differences between virulent and avirulent strains isolated from diseased farmed Senegalese sole based on the protein patterns of their ECP. However, a clear relationship between virulence and presence of prophages has not been established. The differences observed between virulent and nonvirulent strains could be used to design prophylactic strategies against diseases caused by V. harveyi in farmed Senegalese sole.


False-positive results of screening for antibodies to human immunodeficiency virus in chronic hemodialysis patients.
Eighty-three chronic hemodialysis patients were tested for human immunodeficiency virus (HIV) infection. Testing included screening enzyme immunoassay (EIA) for HIV antibodies, competitive EIA for envelope and core antibodies, EIA for HIV antigen, and lymphocyte culture. Five (6%) of the patients had positive screening EIA at low reactivity. Four of these five had antibodies to H-9 cellular antigens. Comparison of the five seropositive patients to matched controls showed no significant differences in number of lymphocytes or helper/suppressor ratio. Six months later, the five patients had negative screening EIA results using a kit with a manufacturing change approved by the Food and Drug Administration that provided improved specificity. In addition, their Western blot analysis was negative. We conclude that (1) false-positive screening EIA results are more common in chronic hemodialysis patients than other populations; (2) evaluation of chronic hemodialysis patients for HIV infection requires confirmatory tests; and (3) newer EIA screening kits appear to have improved specificity.


Urinary exosomes: Emerging therapy delivery tools and biomarkers for urinary system diseases.
Urinary exosomes (UE) are small circular membranous vesicles with a lipid bilayer with a diameter of 40-160 nm secreted by epithelial cells of the kidney and genitourinary system, which can reflect the physiological and functional status of secretory cells. Protein and RNA in exosomes can be used as markers for diseases diagnosis. Urine specimens are available and non-invasive. The protein and RNA in UE are more stable than the soluble protein and RNA in urine, which have broad application prospects in the diagnosis of urinary system diseases. This article reviews the recent advances in the application of protein or RNA in UE as markers to the diagnosis of urinary system diseases.


Rectal cancer in the elderly: patients' perception of bowel control after restorative surgery.
The risk of incontinence is perceived as a relative contraindication to a sphincter-saving procedure in elderly patients with rectal cancer. To investigate this, we reviewed functional results one year after restorative surgery in patients older than aged 75 years. A total of 186 patients older than aged 75 years were diagnosed with rectal cancer during the study period. In 19 patients, no surgery was undertaken, and another 3 patients had a defunctioning stoma only. A local excision was performed in six patients. Of the remaining 158 having a major resection, 133 patients (84 percent) had a restorative anterior resection. The one-year, follow-up records of these patients were studied with particular reference to the patients' perception of bowel function and continence. At one year, 99 patients who had had an anterior resection were alive. In 92 patients, a stoma had been closed or avoided altogether, and these patients formed the study group. Significant problems with bowel function or continence were denied by 78 of 92 patients (85 percent). One patient had already reported severe difficulties and been given a definitive stoma for incontinence. The remaining 14 percent, although experiencing some problems with continence, did not consider the situation serious enough to contemplate a stoma. An elderly patient undergoing anterior resection for rectal cancer has a reasonable expectation of acceptable continence. Age alone should not be a contraindication to a restorative resection.


Large tetraalkyl ammonium cations produce a reduced conductance state in the sheep cardiac sarcoplasmic reticulum Ca(2+)-release channel.
The purified Ca(2+)-release/ryanodine receptor channel of the sheep cardiac muscle sarcoplasmic reticulum (SR) functions as a calcium-activated cation-selective channel under voltage clamp conditions following reconstitution into planar phospholipid bilayers. We have investigated the effect of large tetraalkyl ammonium (TAA) cations, (CnH2n+1)4N+ (n = 4 and 5) on monovalent cation conduction. These cations modify the conductance of the receptor channel at positive holding potentials from the cytosolic side of the channel. Under these conditions, openings are resolved as a mixture of normal full amplitude events and events of reduced conductance. The amplitude of the reduced conductance state is a fixed proportion of the normal open state. As a proportion of all open events, the occurrence of the tetrabutyl ammonium (TBA+) related subconductance state increases with concentration and increasingly positive holding potential. The TBA+ related subconductance state displays similar conduction properties to the unmodified channel; with a linear current-voltage relationship, a similar affinity for K+ and voltage-dependent block by TEA+. A method was used to quantify the voltage dependence of the occurrence of the TBA+ effect, which yielded an effective gating charge of 1.66. A second method based on kinetic analysis of the voltage dependence of transitions between the full open state and the TBA+ related subconductance state produced a similar value. In addition, this analysis revealed that the bulk of the voltage-dependence resided in the off rate. TBA+ related subconductance events, expressed as a proportion of all open events, saturated with increasing TBA+ concentration. Kinetic analysis revealed that this could be entirely accounted for by changes in the on rate. Tetrapentyl ammonium (TPeA+) causes a qualitatively similar effect with a subconductance state of lower amplitude. The voltage-dependence of the effect was comparable to that displayed by TBA+. These findings are interpreted as a form of partial block in which more than one large TAA cation binds at the extremity of the voltage drop to produce an electrostatic barrier for ion translocation.


[Brain imaging in early onset anorexia].
Structural and functional brain alterations in the structures involved in taste processing, emotions regulation and the reward system have been described in anorexia nervosa. The neurodevelopmental origin of this disorder has been recently discussed. In this article, brain-imaging data in early onset anorexia nervosa will be recalled and the relationship between clinical symptoms, normal brain maturation and brain imaging data in adolescents and adults will be discussed.


The sonopartogram.
The assessment of labor progress from digital vaginal examination has remained largely unchanged for at least a century, despite the current major advances in maternal and perinatal care. Although inconsistently reproducible, the findings from digital vaginal examination are customarily plotted manually on a partogram, which is composed of a graphical representation of labor, together with maternal and fetal observations. The partogram has been developed to aid recognition of failure to labor progress and guide management-specific obstetrical intervention. In the last decade, the use of ultrasound in the delivery room has increased with the advent of more powerful, portable ultrasound machines that have become more readily available for use. Although ultrasound in intrapartum practice is predominantly used for acute management, an ultrasound-based partogram, a sonopartogram, might represent an objective tool for the graphical representation of labor. Demonstrating greater accuracy for fetal head position and more objectivity in the assessment of fetal head station, it could be considered complementary to traditional clinical assessment. The development of the sonopartogram concept would require further undertaking of serial measurements. Advocates of ultrasound will concede that its use has yet to demonstrate a difference in obstetrical and neonatal morbidity in the context of the management of labor and delivery. Taking a step beyond the descriptive graphical representation of labor progress is the question of whether a specific combination of clinical and demographic parameters might be used to inform knowledge of labor outcomes. Intrapartum cesarean deliveries and deliveries assisted by forceps and vacuum are all associated with a heightened risk of maternal and perinatal adverse outcomes. Although these outcomes cannot be precisely predicted, many known risk factors exist. Malposition and high station of the fetal head, short maternal stature, and other factors, such as caput succedaneum, are all implicated in operative delivery; however, the contribution of individual parameters based on clinical and ultrasound assessments has not been quantified. Individualized risk prediction models, including maternal characteristics and ultrasound findings, are increasingly used in women's health-for example, in preeclampsia or trisomy screening. Similarly, intrapartum cesarean delivery models have been developed with good prognostic ability in specifically selected populations. For intrapartum ultrasound to be of prognostic value, robust, externally validated prediction models for labor outcome would inform delivery management and allow shared decision-making with parents.


Studying actin-dependent processes in tissue culture.
The cytoskeletal organization of cells that are grown in tissue culture is often very different from that of cells in living organisms. This casts some doubt as to whether information that comes from studying actin-dependent cellular processes--such as cell motility or differentiation--in cells that are cultured under these conditions is physiologically relevant. Studies on cells grown in improved two-dimensional- and three-dimensional-culture systems that closely mimic the in vivo extracellular-matrix environment should provide a more accurate picture of actin-cytoskeletal function in the living organism.


[Muscular absorption of low frequency resonance in animal experiments (author's transl)].
For the further clarification of helicopter pilot's spinal troubles caused by vibration the muscular absorption was examined with the help of animal experiments. On a swinging-table acceleration values were measured in Pirbright cavies; at first this was done under a defence reaction of the muscles, and in second experiment under maximum muscular relaxation. In the tense animal the resonance occurring at 5 Hz was almost completely absorbed, whereas in the relaxed animal a clear resonance--curve could be developed at 5 Hz. On the other hand vibration at a higher frequency up to 15 Hz were to an increasing extent better absorbed in the relaxed animal than in animal with a muscular defence reaction. For this reason the protective function of a not fatigued musculature for the defence of especially badly tolerated resonance vibrations is being discussed.


The vascular endothelial growth factors and receptors family: Up to now the only target for anti-angiogenesis therapy.
Angiogenesis is a complex biological phenomenon essential for a correct embryonic development and for post-natal growth. In adult life, it is a tightly regulated process but in several pathological conditions, angiogenesis results abnormal with either excessive or insufficient proliferation of blood vessels. The pro-angiogenic members of VEGF family, VEGF-A, VEGF-B and placental growth factor (PlGF), and the related receptors, VEGFR-1 and VEGFR-2, have a central and decisive role in pathological angiogenesis. Indeed, they are the targets for anti-angiogenic drugs currently approved: bevacizumab and ranibizumab, that specifically inhibit VEGF-A; aflibercept, that is able to prevent the activity of VEGF-A, VEGF-B and PlGF; several multirtarget tyrosine kinase inhibitors that are able to prevent VEGFR-1 and/or VEGFR-2 signaling. The anti-angiogenesis therapy has represented one of the most active fields of drug discovery of last decade and promises to be further expanded due the wide number of diseases for which it may by applied.


Viral variants that initiate and drive maturation of V1V2-directed HIV-1 broadly neutralizing antibodies.
The elicitation of broadly neutralizing antibodies (bNAbs) is likely to be essential for a preventative HIV-1 vaccine, but this has not yet been achieved by immunization. In contrast, some HIV-1-infected individuals naturally mount bNAb responses during chronic infection, suggesting that years of maturation may be required for neutralization breadth. Recent studies have shown that viral diversification precedes the emergence of bNAbs, but the significance of this observation is unknown. Here we delineate the key viral events that drove neutralization breadth within the CAP256-VRC26 family of 33 monoclonal antibodies (mAbs) isolated from a superinfected individual. First, we identified minority viral variants, termed bNAb-initiating envelopes, that were distinct from both of the transmitted/founder (T/F) viruses and that efficiently engaged the bNAb precursor. Second, deep sequencing revealed a pool of diverse epitope variants (immunotypes) that were preferentially neutralized by broader members of the antibody lineage. In contrast, a 'dead-end' antibody sublineage unable to neutralize these immunotypes showed limited evolution and failed to develop breadth. Thus, early viral escape at key antibody-virus contact sites selects for antibody sublineages that can tolerate these changes, thereby providing a mechanism for the generation of neutralization breadth within a developing antibody lineage.


A prospective study of infections in burn patients.
In a 3-year prospective study, all infections presenting in the burns unit of a university hospital were registered in a specially designed database. Two-hundred and thirty adult patients were included. Eighty-three patients had in all 176 infections, giving an infection rate of 48 per 1000 patient days including both nosocomial and community-acquired infections. Thirty-five blood-stream infections (BSI) occurred in 22 patients; most common micro-organisms were coagulase-negative staphylococci and methicillin-sensitive Staphylococcus aureus. The device-specific BSI rate was 6 per 1000 central venous catheter days, which is low compared to other burn units. The pneumonia rate was 41 per 1000 ventilator days. Seventy-two patients had 107 burn wound infections. Antibiotics were given to only 50% of the burn patients, including 96% of the patients with infection and 26% of those without infection. Most frequently used antimicrobials were cloxacillin, penicillin and gentamicin. The antibiotic resistance rates were low, and multi-resistant bacteria or fungi were rare. The database can be used to evaluate the effects of changes in burn treatment, staffing and design of burn units, and antimicrobial resistance development in relation to antibiotic usage.


Non-union of the scaphoid: early and late management.
The natural history of the fractured scaphoid and its non-union are incompletely understood. Papers are not easily compared without a system of classification. Avascular necrosis is rarely a problem. Unstable fractures should be fixed early to avoid established non-union. Treatment of non-union by graft alone requires prolonged immobilization. Non-union is well treated by grafting and internal fixation. Carpal collapse should be sought and corrected.


Cerebellar ataxia and acute motor axonal neuropathy associated with Anti GD1b and Anti GM1 antibodies.
The anti-GD1b antibody is known to bind to the cerebellar granular layer or spinocerebellar Ia fibers. A few cases of anti-GD1b positive acute inflammatory demyelinating polyneuropathy with prominent cerebellar ataxia were reported. Recently, we encountered a middle-aged woman with Guillain Barré syndrome (GBS) with severe cerebellar ataxia and relatively mild motor weakness. Anti-GD1b Ig G antibody and anti-GM1 Ig G antibody titers were markedly elevated in her serum. She was diagnosed with acute motor axonal neuropathy (AMAN) with prominent cerebellar ataxia based on the results of the serial nerve conduction study suggesting axonal neuropathy. This case presents the clinico-pathogenic role of autoantibodies to the GD1b and the GM1 in acute inflammatory neuropathy.


Persistent cemento-ossifying fibroma of the mandible: report of a case and review of literature.
This case report describes an unusual recurrence of a cemento-ossifying fibroma and illustrates the diagnostic difficulties that may be encountered when treating fibro-osseous lesions.


Mental health and related influencing factors among rural elderly in 14 poverty state counties of Chongqing, Southwest China: a cross-sectional study.
China has the largest elderly population in the world; little attention has been paid to the mental health of elderly in areas of extreme poverty. This is the first study to investigate the mental health of the rural elderly in poverty state counties in Chongqing and was part of the Chongqing 2018 health literacy promotion project. In 2019, a cross-sectional study was conducted to investigate the mental health status of the rural elderly in fourteen poverty state counties of Chongqing, in which a total of 1400 elderly aged ≥ 65 years were interviewed, where mental health status was measured by the ten-item Kessler10 (K10) scale. Ordered multivariate logistic regression was performed to evaluate the influencing factors related to mental health of the elderly in these areas. The average score of K10 in 14 poverty state counties was 17.40 ± 6.31, 47.6% was labeled as good, 30.2% was moderate, 17.0% was poor, and lastly 5.1% was bad, and the mental health status of the elderly in the northeastern wing of Chongqing was better than the one in the southeastern wing of Chongqing. A worse self-rated health was the risk factor for mental health both in the northeastern and southeastern wings of Chongqing (all P < 0.001). Lower education level (OR (95% CI) = 1.45 (1.12-1.87), P = 0.004) was a risk factor in the northeastern wing, whereas older age (OR (95% CI) = 1.33 (1.13-1.56), P = 0.001) was a risk factors in the southeastern wing. The results showed that mental health of the elderly in poverty state counties was poor, especially in the southeastern wing of Chongqing. Particular attention needs to be paid to the males who were less educated, older, and single; female with lower annual per capital income; and especially the elderly with poor self-rated health.


Survivin expressions in human hepatoma HepG2 cells exposed to ionizing radiation of different LET.
Survivin is a member of the inhibitors of apoptosis (IAP) protein family that interferes with post-mitochondrial events including activation of caspases. To examine the regulation of survivin expression in response to irradiation with different linear energy transfer (LET), human hepatoma HepG2 cells were irradiated in vitro with X-rays and carbon ions. Cellular sensitivities to low- and high-LET radiation were determined by colony formation. Survivin expression at mRNA and protein level were measured with RT-PCR and Western blot analyses, respectively. Radiation-induced cell cycle arrest and apoptosis were investigated with flow cytometry. We found that low-LET X-rays induced dose-dependent increases in survivin expression. After exposure to high-LET carbon ions, survivin expression gradually increased from 0 to 4 Gy, and then declined at 6 Gy. More pronounced survivin expression, stronger G(2)/M phase arrest was observed after exposure to carbon ions in comparison with X-rays at doses from 0 to 4 Gy. These observations indicate that there is a differential survivin expression in response to different LET radiations and the cycle arrest mechanism may be associated with it. In addition, our data on induction of apoptosis are compatible with the assumption that survivin expression induced by low-LET X-rays radiation may play a critical role in inhibiting apoptosis. However, after irradiation with ions, an anti-apoptotic function of survivin is not evident, possibly because of the serious damage produced by densely ionizing radiation.


Viral causes of sensorineural hearing loss.
Many viruses, even the common cold, are capable of producing sensorineural hearing loss. Treatment so far available is ineffective in correcting these losses. However, as the technology of vaccination advances, it is hoped that these can be prevented. Poliomyelitis and smallpox have been virtually eliminated from this country. Measles, rubella, and mumps are coming under control. Perhaps in the future we will be able to completely control influenza and the common cold and thus prevent many of the sensorineural hearing losses that we see today.


The decline in transurethral resection of the prostate gland in Irish public hospitals between 2005 and 2021.
Lower urinary tract symptoms due to an enlarged prostate is a common condition. Transurethral resection of the prostate gland (TURP) has been the gold standard treatment. The objective of this study was to assess the trends in the prevalence of TURP procedures in Irish public hospitals within the period of 2005-2021. In addition, we explore the attitudes and practices of urologist in Ireland on this topic. An analysis using the Hospital In-Patient Enquiry (HIPE) system using code 37203-00 was undertaken. 16,176 discharges contained the code of interest and had undergone a TURP procedure. The data from this cohort was further analysed. In addition, members of the Irish Society of Urology undertook a bespoke questionnaire to understand the TURP surgery practices. There has been a substantial decline in the prevalence of TURP procedures in Irish public hospitals from 2005 to 2021. The number of patients discharged from Irish hospitals with a TURP procedure was 66% less in 2021 compared to 2005. 75% (n = 36) of urologist surveyed felt that the declining TURP numbers were due to lack of resources, access to theatre/inpatient beds and outsourcing. 91.5% (n = 43) felt that the declining TURP numbers would result in a lack of training opportunities for trainees, 83% (39) felt this has increased morbidity for patients. TURP procedures in Irish public hospitals has declined over the 16-year period studied. This decline is a concern for patient morbidity and urology training.


RBM24 is localized to stress granules in cells under various stress conditions.
Stress granules (SGs) are formed when untranslated messenger ribonucleoproteins (mRNPs) accumulate in cells under stress, and are thought to minimize stress-induced damage and promote cell survival. RBM24 (RNA-binding motif protein 24) is an RNA-binding protein that plays pivotal roles in regulating the stability or translation initiation of target mRNAs as well as alternative splicing of target pre-mRNAs. Its important physiological functions are highlighted by the fact that Rbm24 knockout mice or zebrafish suffer from dysfunction of heart, eye, and inner ear. Here we show that RBM24 is recruited into SGs under various stress conditions, suggesting that it might protect its target RNAs in cells under stress. However, SG formation is unaffected when Rbm24 expression is down-regulated. Nevertheless, RBM24 overexpression in cultured cells is sufficient to induce SG formation, suggesting that RBM24 might play an important role in SG formation. In conclusion, our present work reveals that RBM24 is a SG component, which implies that RBM24 could protect its target mRNAs in stressed cells.


Potential of butanol production from Thailand marine macroalgae using Clostridium beijerinckii ATCC 10132-based ABE fermentation.
The economical bio-butanol-based fermentation process is mainly limited by the high price of first-generation biomass, which is an intensive cost for the pretreatment of second-generation biomass. As third-generation biomass, marine macroalgae could be potentially advantageous for conversion to clean and renewable bio-butanol through acetone-butanol-ethanol (ABE) fermentation. In this study, butanol production from three macroalgae species (Gracilaria tenuistipitata, Ulva intestinalis, and Rhizoclonium sp.) by Clostridium beijerinckii ATCC 10132 was assessed comparatively. The enriched C beijerinckii ATCC 10132 inoculum produced a high butanol concentration of 14.07 g L-1 using 60 g L-1 of glucose. Among the three marine seaweed species, G. tenuistipitata exhibited the highest potential for butanol production (1.38 g L-1 ). Under the 16 conditions designed using the Taguchi method for low-temperature hydrothermal pretreatment (HTP) of G. tenuistipitata, the maximum reducing sugar yield rate of 57.6% and ABE yield of 19.87% were achieved at a solid to liquid (S/L) ratio of 120, temperature of 110°C, and holding time of 10 min (Severity factor, R0 1.29). In addition, pretreated G. tenuistipitata could be converted to 3.1 g L-1 of butanol using low-HTP at an S/L ratio of 50 g L-1 , temperature of 80°C (R0 0.11), and holding time of 5 min.


Genetic diversity of the DBLalpha region in Plasmodium falciparum var genes among Asia-Pacific isolates.
In Plasmodium falciparum a highly polymorphic multi-copy gene family, var, encodes the variant surface antigen P. falciparum erythrocyte membrane protein 1 (PfEMP1), which has an important role in cytoadherence and immune evasion. Using previously described universal PCR primers for the first Duffy binding-like domain (DBLalpha) of var we analysed the DBLalpha repertoires of Dd2 (originally from Thailand) and eight isolates from the Solomon Islands (n=4), Philippines (n=2), Papua New Guinea (n=1) and Africa (n=1). We found 15-32 unique DBLalpha sequence types among these isolates and estimated detectable DBLalpha repertoire sizes ranging from 33-38 to 52-57 copies per genome. Our data suggest that var gene repertoires generally consist of 40-50 copies per genome. Eighteen DBLalpha sequences appeared in more than one Asia-Pacific isolate with the number of sequences shared between any two isolates ranging from 0 to 6 (mean=2.0 +/-1.6). At the amino acid level DBLalpha sequence similarity within isolates ranged from 45.2 +/- 7.1 to 50.2 +/- 6.9%, and was not significantly different from the DBLalpha amino acid sequence similarity among isolates (P>0.1). Comparisons with published sequences also revealed little overlap among DBLalpha sequences from different regions. High DBLalpha sequence diversity and minimal overlap among these isolates suggest that the global var gene repertoire is immense, and may potentially be selected for by the host's protective immune response to the var gene products, PfEMP1.


Nonpuerperal breast infection.
We undertook a microbiological study of purulent specimens from women with symptomatic breast abscesses. Fifty-one purulent samples were collected in 2 periods (December 1991-April 1992 and January 1994-June 1994) from nonpuerperal breast abscesses in 44 patients attending our hospital. One of the most frequently isolated microorganisms was Proteus mirabilis (9 patients, 20.4%), present as a pure culture in all but 1 specimen (isolated together with Peptostreptococcus spp.). Staphylococcus aureus was isolated in 10 specimens, 6 of which were post-tumorectomy abscesses. Polymicrobial anaerobic flora were isolated in 11 specimens (21.5%); Staphylococcus epidermidis in 4 (8%); and Streptococcus milleri,Alcaligenes sp., and mixed aerobic-anaerobic flora in 1 specimen each. The 7 remaining samples (13.7%) were negative bacteriological cultures. We draw attention to the frequent isolation of P. mirabilis in recurrent and torpid breast abscesses in 4 women in whom surgery was necessary in addition to antibiotic treatment.


Caudal regression with cake kidney and a single ureter: a case report.
We report the second case of a cake kidney with a single ureter accompanied by caudal regression. The embryology, presentation and management of this malformation are delineated.


Changes in KCNQ2 immunoreactivity in the amygdala in two rat models of temporal lobe epilepsy.
Potassium channels containing the KCNQ2 subunit play an important role in the regulation of neuronal excitability and therefore have been implicated in epilepsy. This study describes the expression of KCNQ2 subunit immunoreactivity in the basolateral amygdala in two rat models of temporal lobe epilepsy, (1) amygdala kindling and (2) spontaneously epileptic rats after status epilepticus induced by hippocampal electrical stimulation. KCNQ2 subunit immunoreactivity was assessed with a commercial antibody raised against a C-terminal part of the KCNQ2 protein. We show that KCNQ2 subunit immunoreactivity is upregulated in the basolateral amygdala in both models and that generalized seizures are required to induce this upregulation. We hypothesize that the upregulation of potassium channels containing the KCNQ2 subunit might represent a mechanism to counteract seizures in experimental temporal lobe epilepsy.


Cell cycle phase-specific cDNA libraries reflecting phase-specific gene expression of Ehrlich ascites cells growing in vivo.
Asynchronous populations of Ehrlich ascites tumor cells grown in vivo were separated by centrifugal elutriation into fractions of G1-, S-, and G2/M-phase cells with less than 10% cross-contamination. Cytoplasmic mRNA from phase-synchronous cells was used to prepare cDNA which was ligated with bacteriophage lambda gt10 arms and amplified in Escherichia coli C600 hfl-. EcoRI digests of DNA isolated from the sublibraries (G1, S, G2/M) were submitted to Southern hybridizations with radiolabeled probes either (a) for genes whose phase-specific expression is clearly documented, thymidine kinase, dihydrofolate reductase, and thymidylate synthase, or (b) for genes whose change of expression during the cell cycle is likely, lamin C, beta-actin, alpha- and beta-tubulin, c-myc, c-fos, p53. The cDNA sequences for genes of group (a) were found to be significantly enriched in DNA of the S-phase library indicating that the cell cycle phase-specific patterns of the respective mRNA levels are conserved in the sublibraries. Sequences belonging to group (b) were also found to be enriched in DNA isolated from the sublibraries: c-fos in G1 phase, lamin C, beta-actin, tubulins, c-myc in S phase, and p53 in G1/S phase. The unexpected prevalence of c-myc and alpha-tubulin in the S-phase library is supported by Northern analysis of RNA from phase-synchronous cells. Non-phase-specific, randomly chosen sequences hybridized equally strong with DNA isolated from the different sublibraries. No significant changes of the patterns of hybridization signals were observed with DNA from different amplifications of the sublibraries when analyzed with the same DNA probe indicating that the cDNA complexities are well conserved during amplifications. Consequently, the sublibraries are useful to obtain information about the cell cycle phase-specific expression of mRNAs for other genes of interest. Since the sublibraries reflect mRNA levels of the cells growing in vivo they supply data on the physiological in vivo pattern of gene expression undisturbed by potentially unphysiological in vitro conditions.


Three complementation groups in Cockayne syndrome.
After 16 Jm-2 of UV-irradiation non-dividing normal cells recover normal rates of RNA synthesis within 24 h, whereas in cells from donors with Cockayne syndrome (CS) the rate of RNA synthesis gradually declines. Cultures of a mixed population from 2 CS donors were fused with polyethylene glycol; subsequently they were UV-irradiated and RNA synthesis was measured autoradiographically in mono-, bi-, and multinuclear cells. Genetic complementation was indicated by high levels of RNA synthesis in bi- and multinuclear cells when compared with mononuclear cells. Using this assay, 11 CS strains have been assigned to three complementation groups: 2 into group A, 8 into group B and 1 into group C. The strain in group C is derived from an individual who also had xeroderma pigmentosum (XP), and was the sole known representative of XP-complementation group B.


Sedentary behavior and the risk of stroke: A systematic review and dose-response meta-analysis.
The sedentary behavior in people's daily life has continued to increase in recent years, causing many studies to focus on its relationship with diseases. Several studies have shown that sedentary behavior is an independent risk factor for cardiovascular disease and metabolic disease. Therefore, we performed a meta-analysis to assess the association between sedentary behavior and the risk of stroke. Two independent investigators searched for prospective cohort studies on the association between sedentary behavior and stroke risk, published before February 2022. We pooled adjusted effect size and performed the dose-response analysis by random-effect model. Seven studies with 677,614 participants and 15,135 stroke events during a median follow-up of 12.2 years were included. The pooled hazard ratio (HR) of stroke was 1.16 (95% confidence interval [CI]: 1.09-1.24) with no significant heterogeneity (I2 = 0.0%, p for heterogeneity = 0.983). In dose-response analysis, a nonlinear association between sedentary behavior and stroke risk was discovered. Stroke risk began to increase when sedentary time exceeded 3.7 h/d (HR, 1.01; 95% CI, 0.97-1.05). And when reached 11 h/d, a significantly increased risk of stroke was observed (HR, 1.21; 95% CI 1.12-1.31). A nonlinear association was found in the dose-response analysis, with increased risk only when sedentary time exceeded a certain level. Further research is needed to explain the biological mechanisms by which sedentary time above a certain threshold significantly increases stroke risk. (PROSPERO registration number: CRD42022311544).


Cost drivers in patient-controlled epidural analgesia for postoperative pain management after major surgery.
In this retrospective study, we determined efficiency, treatment length, and resource use for postoperative pain management with patient-controlled epidural analgesia (PCEA) in 350 consecutive patients undergoing major abdominal, thoracic, gynecological, or orthopedic surgery. Average pain scores on a visual analog scale were 16 +/- 23 and 9 +/- 16 (visual analog scale range, 0 to 100) on postoperative Days 1 and 3, respectively, and were similar among groups. The treatment length was 4.9 +/- 2.2 days in general surgical, 5.2 +/- 3.1 days in gynecological, and 4.5 +/- 2.8 days in orthopedic patients. The total volumes of the mixture of local anesthetic and opioid received epidurally were 707 +/- 507 mL, 770 +/- 576 mL, and 593 +/- 456 mL in the general surgical, gynecological, and orthopedic groups, respectively. The average total costs for all groups for the full treatment course with PCEA were 447 +/- 218 per case (1 equals approximately US dollar 1). Fifty-one percent of these costs were staff costs, 20% were costs for the applied drugs, 15% were costs for PCEA pumps and pump material, and 13% were costs for the initial catheter insertion. In the light of these costs and the availability of less costly alternatives, measurements for cost containment by using PCEA are recommended. Because treatment length is the main cost driver both for drug and staff costs, close monitoring of treatment length and a predefined migration path to alternative techniques after PCEA should be considered. Patient-controlled epidural analgesia is increasingly used as first-line treatment for postoperative pain management. In this study, costs and cost drivers are analyzed for the first time for this new technique, based on 350 cases of pain therapy after major surgery in a university hospital.


Recruitment of Nox4 to a plasma membrane scaffold is required for localized reactive oxygen species generation and sustained Src activation in response to insulin-like growth factor-I.
Nox4-derived ROS is increased in response to hyperglycemia and is required for IGF-I-stimulated Src activation. This study was undertaken to determine the mechanism by which Nox4 mediates sustained Src activation. IGF-I stimulated sustained Src activation, which occurred primarily on the SHPS-1 scaffold protein. In vitro oxidation experiments indicated that Nox4-derived ROS was able to oxidize Src when they are in close proximity, and Src oxidation leads to its activation. Therefore we hypothesized that Nox4 recruitment to the plasma membrane scaffold SHPS-1 allowed localized ROS generation to mediate sustained Src oxidation and activation. To determine the mechanism of Nox4 recruitment, we analyzed the role of Grb2, a component of the SHPS-1 signaling complex. We determined that Nox4 Tyr-491 was phosphorylated after IGF-I stimulation and was responsible for Nox4 binding to the SH2 domain of Grb2. Overexpression of a Nox4 mutant, Y491F, prevented Nox4/Grb2 association. Importantly, it also prevented Nox4 recruitment to SHPS-1. The role of Grb2 was confirmed using a Pyk2 Y881F mutant, which blocked Grb2 recruitment to SHPS-1. Cells expressing this mutant had impaired Nox4 recruitment to SHPS-1. IGF-I-stimulated downstream signaling and biological actions were also significantly impaired in Nox4 Y491F-overexpressing cells. Disruption of Nox4 recruitment to SHPS-1 in aorta from diabetic mice inhibited IGF-I-stimulated Src oxidation and activation as well as cell proliferation. These findings provide insight into the mechanism by which localized Nox4-derived ROS regulates the sustained activity of a tyrosine kinase that is critical for mediating signal transduction and biological actions.


Acute exercise impacts AhR and PD-1 levels of CD8+ T-cells-Exploratory results from a randomized cross-over trial comparing endurance versus resistance exercise.
The programmed cell death protein 1 (PD-1) has become a promising target in cancer immunotherapy. PD-1 expression of CD8+ T-cells may be increased via the exploitation of aryl hydrocarbon receptor (AhR) signaling with kynurenine (KYN) as a ligand. Since exercise affects KYN metabolism, we exploratory investigated the influence of acute exercise bouts on AhR and PD-1 levels of CD8+ T-cells. In this study, 24 healthy males (age: 24.6 ± 3.9 years; weight 83.9 ± 10.5 kg; height: 182.4 ± 6.2 cm) completed a single bout of endurance (EE) and resistance exercise (RE) in a randomly assigned order on separate days. Blood samples were drawn before (t0), after (t1), and 1 h after (t2) both conditions. T-cell populations, the level of cytoplasmic AhR, and surface PD-1 were assessed by flow cytometry. T-cell populations changed over time, indicated by an increase in the absolute numbers of CD3+ lymphocytes after EE (p < .001) and RE (p = .036) and in PD-1+ CD8+ T-cells after EE (p = .021). Proportions of T-cell populations changed only after EE (t0-t2: p = .029; t1-t2: p = .006). The level of cytoplasmic AhR decreased immediately after exercise in both exercise conditions (EE: p = .009; RE: p = .036). The level of surface PD-1 decreased 1 h after EE (p = .005). We analyzed the level of surface PD-1 and cytoplasmic AhR following acute physical exercise for the first time. Especially EE was observed to impact both AhR and PD-1 levels, undermining its role as the AhR-PD-1 axis modulator. These results provide new insights into the impact of exercise on AhR-signaling, which could potentially be relevant for various chronic diseases.


Synthetic surfactant with a recombinant surfactant protein C analogue improves lung function and attenuates inflammation in a model of acute respiratory distress syndrome in adult rabbits.
In acute respiratory distress syndrome (ARDS) damaged alveolar epithelium, leakage of plasma proteins into the alveolar space and inactivation of pulmonary surfactant lead to respiratory dysfunction. Lung function could potentially be restored with exogenous surfactant therapy, but clinical trials have so far been disappointing. These negative results may be explained by inactivation and/or too low doses of the administered surfactant. Surfactant based on a recombinant surfactant protein C analogue (rSP-C33Leu) is easy to produce and in this study we compared its effects on lung function and inflammation with a commercial surfactant preparation in an adult rabbit model of ARDS. ARDS was induced in adult New Zealand rabbits by mild lung-lavages followed by injurious ventilation (VT 20 m/kg body weight) until P/F ratio < 26.7 kPa. The animals were treated with two intratracheal boluses of 2.5 mL/kg of 2% rSP-C33Leu in DPPC/egg PC/POPG, 50:40:10 or poractant alfa (Curosurf®), both surfactants containing 80 mg phospholipids/mL, or air as control. The animals were subsequently ventilated (VT 8-9 m/kg body weight) for an additional 3 h and lung function parameters were recorded. Histological appearance of the lungs, degree of lung oedema and levels of the cytokines TNFα IL-6 and IL-8 in lung homogenates were evaluated. Both surfactant preparations improved lung function vs. the control group and also reduced inflammation scores, production of pro-inflammatory cytokines, and formation of lung oedema to similar degrees. Poractant alfa improved compliance at 1 h, P/F ratio and PaO2 at 1.5 h compared to rSP-C33Leu surfactant. This study indicates that treatment of experimental ARDS with synthetic lung surfactant based on rSP-C33Leu improves lung function and attenuates inflammation.


Surface-enhanced Raman scattering and density functional theory study of 1,4-benzenedithiol and its silver complexes.
This paper experimentally and theoretically investigated Raman and surface-enhanced Raman scattering (SERS) of 1,4-benzenedithiol (1,4-BDT). Density functional theory methods were used to study Raman scattering spectra of isolated 1,4-BDT and 1,4-BDT-Agn (n=2,4,6) complexes with B3LYP/6-311+g(d)(C,H,S)/Lanl2dz(Ag) basis set. A full assignment of the Raman spectrum of 1,4-BDT has been made based on the DFT analysis. The calculated data showed good agreement with experimental observations. The adsorption sites, metal cluster size, and HOMO-LUMO energies are discussed to give insight in the SERS mechanisms for 1,4-BDT molecules.


The inhibition of lipopolysaccharide-induced macrophage inflammation by 4 compounds in Hypericum perforatum extract is partially dependent on the activation of SOCS3.
Our previous studies found that 4 compounds, namely pseudohypericin, amentoflavone, quercetin, and chlorogenic acid, in Hypericum perforatum ethanol extract synergistically inhibited lipopolysaccharide (LPS)-induced macrophage production of prostaglandin E2 (PGE2). Microarray studies led us to hypothesize that these compounds inhibited PGE2 production by activating suppressor of cytokine signaling 3 (SOCS3). In the current study, siRNA was used to knockdown expression of SOCS3 in RAW 264.7 macrophages and investigated the impact of H. perforatum extract and the 4 compounds on inflammatory mediators and cytokines. It was found that the SOCS3 knockdown significantly compromised the inhibition of PGE2 and nitric oxide (NO) by the 4 compounds, but not by the extract. The 4 compounds, but not the extract, decreased interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), while both lowered interleukine-1β. SOCS3 knockdown further decreased IL-6 and TNF-α. Pseudohypericin was the major contributor to the PGE2 and NO inhibition in cells treated with the 4 compounds, and its activity was lost with the SOCS3 knockdown. Cyclooxygenase-2 (COX-2) and inducible NO synthase protein expression were not altered by the treatments, while COX-2 activity was decreased by the extract and the 4 compounds and increased by SOCS3 knockdown. In summary, it was demonstrated that the 4 compounds inhibited LPS-induced PGE2 and NO through SOCS3 activation. The reduction of PGE2 can be partially attributed to COX-2 enzyme activity, which was significantly elevated with SOCS3 knockdown. At the same time, these results also suggest that constituents in H. perforatum extract were alleviating LPS-induced macrophage response through SOCS3 independent mechanisms.


Reovirus as a successful ex vivo purging modality for multiple myeloma.
Autologous stem cell rescue (ASCT) following high-dose myeloablative chemotherapy is considered to be a therapeutic option for many multiple myeloma (MM) patients; however relapse post ASCT presents a major challenge. The oncolytic potential of reovirus has been previously demonstrated and is currently undergoing phase I monotherapy clinical trials for MM and phase II/III clinical trials for solid tumors. Here we tested the hypothesis that reovirus can successfully purge MM in a murine model that partially recapitulates human MM. RPMI 8226, MM1S, H929 and U266 human myeloma cell lines were exposed to reovirus and oncolysis was assessed. Apheresis product admixed with MM cells was purged with live reovirus (LV) or dead virus (DV) and purging efficacy was monitored via flow cytometry, reverse transcribed-PCR (RT-PCR) and disease relapse in non obese diabetic/severe combined immune deficient (NOD/SCID) mice. Significant LV purging was seen with MM1S, H929 and U266 and the complete ex vivo purging achieved with RPMI 8226 was confirmed by flow cytometry, RT-PCR and absence of disease relapse in vivo. Mice that received LV-purged autografts exhibited 100% survival in comparison to mice that received DV-purged controls. Reovirus's unique ability to kill MM while sparing hematopoietic stem cells places it as an attractive purging agent for MM during ASCT.


The effects of kininase II inhibition by SQ 14225 on kidney kallikrein-kinin and prostaglandin systems in conscious dogs.
The novel kininase II inhibitor SQ 14225 was intravenously administered to normal conscious dogs at the dose of 0.1 mg/kg. Urine kinin excretion increased from 14 +/- 6 ng/h to 163 +/- 18 ng/h. Separation by column chromatography showed that the increased urine kinin excretion was due to increased excretion of bradykinin. The enhanced urinary kinin excretion was associated by increased sodium (70%) and decreased kallikrein (27%) excretion. Urine volume and urinary prostaglandin excretion were not significantly affected by SQ 14225 treatment.


Studies on the immuno-modulating and antitumor activities of Ganoderma lucidum (Reishi) polysaccharides: functional and proteomic analyses of a fucose-containing glycoprotein fraction responsible for the activities.
A fucose-containing glycoprotein fraction which stimulates spleen cell proliferation and cytokine expression has been identified from the water-soluble extract of Ganoderma lucidum. Proteomic analysis of mouse spleen cells treated with this glycoprotein fraction showed approximately 50% change of the proteome. Further studies on the activities of this glycoprotein fraction through selective proteolysis and glycosidic cleavage indicate that a fucose containing polysaccharide fraction is responsible for stimulating the expression of cytokines, especially IL-1, IL-2 and INF-gamma.


The effect of upper cervical or sacroiliac manipulation on hip flexion range of motion.
To compare the effectiveness of an upper cervical manipulation and a manipulation of the sacroiliac joint for increasing hip range of motion. Clinical cohort study. Macquarie University Centre for Chiropractic Outpatient Clinic. Fifty-two randomly chosen university students aged 18 to 34 yr. A reliable hand-held dynamometer was used to determine the end point of range of motion before and after the application of a treatment. Three groups of subjects were created: cervical manipulation, sacroiliac manipulation and sham/placebo. Range of motion of the hip in flexion (SLR) was used as the independent variable. The two manipulative treatments resulted in increased flexion range of motion at the hip. Statistical analysis revealed that only the upper cervical manipulation procedure increased hip flexion range of motion significantly. The results suggest that manual therapy of the neck may affect hip range of motion in normal adults. Findings such as these may indicate the existence of a link between the cervical spine and the lower extremity.


[The influence of elicitor on the distribution pattern of microtubule and the cytosolic calcium in mesophyll protoplast of wheat].
By using immunofluorescence labeling technique and incubating the Fluo-3 AM of Ca2+ fluorescein, wheat (Triticum aestivum) cultivars Lovrin 10 and 5389 were selected as mesophyll protoplast material interaction with elicitor in this paper for studying the relationship between the pattern of microtubule and [Ca2+]cyt. Confocal Laser Scanning Microscopy (CLSM) was used and pharmacologic examination was set. The results indicated: when treated with elicitor, the cytosolic calcium in the protoplasts from the Lovrin 10 was increasing and the pattern of microtubule showed depolymerization. In the protoplasts which MTs were depolymerized by oryzalin prior to elicitor treatment, the increase of cytosolic calcium was even higher than that in the one which was not depolymerized.


Left Ventricular Assist Device Exchange Increases Heart Transplant Wait-List Mortality.
The new heart transplant allocation criteria prioritize inpatients who require temporary mechanical circulatory support and give lower urgency to candidates on a durable left ventricular assist device (LVAD) who require a device exchange. This study explores whether the latter group should warrant higher priority to reduce wait-list mortality. This is a retrospective observational study of 13,113 adult heart transplant candidates in the Organ Procurement and Transplantation Network database who underwent LVAD implantation between 2007 and 2017. It evaluates the impact of LVAD exchange on the composite endpoint of death or removal from the wait list owing to worsening medical condition 1 y after device implantation. There were 1085 pump exchanges in 954 patients (7% of candidates), of which 22% were women. The pump exchange rate was 5.92 events per 100 patient-years. One-year survival was lower for those who required a pump exchange (76.3% versus 88.5%, logrank P < 0.001). This was congruent with the risk-adjusted mortality 1-y after implantation (hazards ratio: 2.56, 95% confidence interval: 2.18-3.00, P < 0.001). These findings indicate that among candidates awaiting heart transplantation with a durable LVAD, undergoing pump exchange doubles the risk of 1-y mortality. Giving priority to these candidates may reduce wait-list mortality.


Regulation of the superoxide-generating NADPH oxidase by a small GTP-binding protein and its stimulatory and inhibitory GDP/GTP exchange proteins.
The superoxide-generating NADPH oxidase system in phagocytes consists of at least membrane-associated cytochrome b558 and three cytosolic components named SOCI/NCF-3/sigma 1/C1, SOCII/NCF-1/p47-phox, and SO-CIII/NCF-2/p67-phox. p47-phox and p67-phox were isolated, and their primary structures were determined, but SOCI has not been well characterized. In the present study, we first purified SOCI to homogeneity from the cytosol fraction of the differentiated HL-60 cells. The purified SOCI was a small GTP-binding protein (G protein) with a M(r) of about 22,000. The guanosine 5'-(3-O-thio)triphosphate-bound form, but not the GDP-bound form, of this small G protein showed the SOCI activity. The partial amino acid sequence of SOCI thus far determined was identical to the amino acid sequence deduced from the cDNA encoding rac2 p21. None of the purified small G proteins, including Ki-ras p21, smg p21B/rap1B p21, rhoA p21, and rac1 p21, showed the SOCI activity. These results indicate that SOCI is a small G protein very similar, if not identical, to rac2 p21. The GDP/GTP exchange reaction of SOCI was stimulated and inhibited by stimulatory and inhibitory GDP/GTP exchange proteins for small G proteins, named smg GDS and rho GDI, respectively. The NADPH oxidase activity was also stimulated and inhibited by smg GDS and rho GDI, respectively. These results indicate that the superoxide-generating NADPH oxidase system is regulated by both smg GDS and rho GDI through rac2 p21 or the rac2-related small G protein in phagocytes.


Lipid profile, oxidant-antioxidant status and glycoprotein components in hyperlipidemic patients with/without diabetes.
Plasma lipoproteins are protected against oxidative modification by the antioxidant defense system. An imbalance in the antioxidant defense system seems to result from the accumulation of low density lipoprotein (or) very low density lipoprotein in the course of hyperlipidemia. The lipid profile, glycoprotein components, glucose, total proteins, albumin, lipid peroxidation and antioxidant status of plasma and erythrocytes were investigated in hyperlipidemic patients and hyperlipidemic patients with diabetes on treatment with glibenclamide (or) glipizide along with normal subjects and compared. A significant increase was observed in the levels of total cholesterol (TC), VLDL-C, triglycerides (TG), lipid peroxidation, ceruloplasmin (CP), glycoprotein components and glucose in the hyperlipidemic patients with/without diabetes and the increase was more pronounced (except TC) in patients with diabetes. The activities of erythrocyte antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and plasma concentrations of vitamins C and E, and reduced glutathione (GSH) decreased in hyperlipidemic patients with/without diabetes. GPx, CAT, vitamin C and GSH levels exhibited a further decrease in hyperlipidemic patients with diabetes. An enhanced levels of VLDL-C, TC/HDL-C ratio, TG, lipid peroxidation, glycoprotein components, and decreased concentrations of total proteins (TPs) and albumin were observed in hyperlipidemic patients with diabetes while the decrease was more marked in GSH, vitamin C, CAT and GPx among antioxidants.