Alterations in body lean angle in lame horses before and after diagnostic analgesia in straight lines in hand and on the lunge.
Altered body lean has been subjectively observed during lungeing in lame horses. The objectives were to quantify the influence of lameness on body lean in trot on the lunge and to investigate the influence of improvement in lameness on the differences in body lean between reins. Thirteen lame horses were trotted in straight lines and lunged on a 10m-diameter circle on both reins before and after lameness was subjectively substantially improved by diagnostic analgesia. A global position system-aided inertial measurement unit attached to the tubera sacrale quantified body lean. Differences between reins in body lean before and after diagnostic analgesia were calculated and means were determined. Five and eight horses had unilateral and bilateral hindlimb lameness, respectively. Two of five horses with unilateral and three of eight horses with bilateral lameness leaned more on the rein with the lame or lamer hindlimb on the inside of the circle (difference between reins 5-8°). Two of five horses with unilateral and two of eight horses with bilateral lameness leaned more on the rein with the lame or lamer hindlimb on the outside of the circle (4-10°). Four horses, one with unilateral and three with bilateral lameness, had only 1° difference in body lean angle between left and right reins. When lameness was improved by diagnostic analgesia, the body lean changed significantly towards similar leaning on left and right reins (mean angle changed from 8.8° to 10.0° (P=0.03) on one rein and 13.4° to 10.8° (P=0.002) on the other rein). It was concluded that body lean becomes more symmetrical between reins after improvement in lameness using diagnostic analgesia.


In vitro activity of dicloxacillin against methicillin-susceptible and methicillin-resistant Staphylococcus-aureus.
A total of 54 strains of Staphylococcus aureus, 37 methicillin-susceptible (MSSA) and 17 methicillin-resistant (MRSA), were investigated for their susceptibility to dicloxacillin as compared to methicillin and oxacillin by agar plate dilution at two different temperatures of incubation (30 degrees C and 37 degrees C). Against MSSA strains we found a slight but significant increase (0-2 dilution steps) in minimal inhibitor concentrations (MICs) for all three antibiotics with decrease in incubation temperature. Against MRSA strains methicillin and oxacillin showed a 3-6 fold increase in median MIC with decrease incubation temperature. For dicloxacillin, in contrast, there were no significant differences in median MICs (i.e. 0.4 mg/l) against MSSA strains at 30 degrees C or MRSA strains at either incubation temperature. Population-analysis of the MRSA strains revealed, however, that a highly dicloxacillin-resistant subpopulation appeared with a frequency of 10(-6) to 10(-7). Such heterogenous resistance of MRSA strains to dicloxacillin probably prohibits the use of dicloxacillin against serious infections caused by these pathogens.


Effect of exogenous lipase on subcutaneous adipose tissue in a porcine animal model.
Topical exogenous lipase has been approved for cosmetic use and has been used to mobilize fat from adipocytes. The objective of this study was to determine the effects of exogenous lipase in the subcutaneous adipose tissue. Three different concentrations of exogenous lipase 1× (2 Units per ml), 5× (10 units per ml), and 10× (20 units per ml) were applied in a porcine model. Normal saline (NS) solution (as negative control) and phosphatidylcholine (as positive control) were also injected. Skin and subcutaneous tissue biopsies, up to the fascia, were obtained from each injection site on the 3rd day after injection. The number of cells per 20× field was counted as an indirect measurement of the size of the adipocytes. For 1× lipase, the number of cells per field was 47.80 (±7.63) versus 27.26 (±4.93), and 34.66 (±6.84) for NS, and phosphatidylcholine, respectively. For 5× lipase, the count was 36.06 (±4.74) versus 24.13 (±5.18), and 33.2 (±9.34). For 10× lipase, it was 40.06 (±4.35) versus 29.26 (±2.34) and 32.66 (±6.30) (p < .05 for all groups). A higher number of cells per field were observed in the lipase samples, inferring a decreased volume of adipocytes. No inflammation and/or loss of cell architecture were evidenced in the exogenous lipase groups.


Rat cytomegalovirus: induction of and sensitivity to interferon.
Rat interferon (IFN) in a dose-dependent fashion inhibited the cytopathic effect of rat cytomegalovirus (RCMV) in rat cell cultures. Treatment of Lewis and Brown Norway (BN) rats with intraperitoneal injections of IFN reduced the amount of virus recovered from the spleens at 3 days post infection (p.i.) and from the salivary glands at 10 and 21 days p.i. In cell cultures, RCMV failed to induce detectable amounts of IFN. Small amounts of IFN were detectable in the serum of BN rats at 3 days p.i. In Lewis rats no IFN was found in serum at any time during the first month p.i.


Investigations on the binding of erythrogenic toxin A of Streptococcus pyogenes on human peripheral blood lymphocytes. II. Identification of toxin-binding lymphocytes and characterization of the receptor.
Gold complexes of erythrogenic toxin A (ETA) of Streptococcus pyogenes (strain NY-5) were bound to human lymphocytes that could be characterized as T cells by rosetting or labelling with OKT 3 antibody and anti-mouse-IgG-gold. Using OKT 4 and OKT 8 antibodies, respectively, it was demonstrated that binding of ETA was confined to the T helper/inducer cell subpopulation. Treatment of lymphocytes with neuraminidase enhanced the binding of ETA-gold, whereas trypsin treatment resulted in its reduction. Gangliosides of type II or type III and D-galactose only slightly decreased the ETA-gold binding. Preincubation of living lymphocytes with the mitogens concanavalin A or the Lens culinaris-lectin enhanced the binding of ETA-gold.


The effect of oral anticoagulant use before visit for patients with COVID-19 on mortality: A meta-analysis.
Anticoagulants have been used as therapeutic or prophylactic agents in COVID-19 and seem to be more beneficial. However, the advantage of oral anticoagulant (OAC) consumption before visit in lowering mortality in COVID-19 patients remains debatable. This meta-analysis aimed to evaluate the effect of OAC use before visit on mortality using the hazard ratio (HR) to estimate the effect of time-to-event endpoints. We conducted a literature search in the PubMed and ProQuest databases for any studies comparing groups consuming OAC to no-OAC before visit for mortality in patients with COVID-19. We calculated the overall HRs and their variances across the studies using the random-effects model to obtain pooled estimates. We included 12 studies which had sample sizes ranging from 70 to 459,402 patients. A meta-analysis comparing OAC therapy and non-OAC consumption in COVID-19 patients before visit revealed no decrease in all-cause mortality (HR = 0.92, 95% confidence interval [CI]: 0.83-1.02, P = 0.12; I2 = 68%). However, subgroup analysis of laboratory-confirmed populations revealed that OAC use before visit had a beneficial effect on mortality (HR = 0.84, 95% CI: 0.73-0.98, P = 0.02; I2 = 56%). The use of OAC before visit had no beneficial effect on all-cause mortality in COVID-19 patients.


The frequency of prescription of immediate-release nifedipine for elderly patients in Germany: utilization analysis of a substance on the PRISCUS list of potentially inappropriate medications.
Immediate-release nifedipine is on the PRISCUS list of drugs that should not be given to elderly patients. We studied the use of this calcium-channel blocker under real-life conditions. In 2009, we carried out a cross-sectional study based on the Statutory Health Insurance Sample AOK Hesse/KV Hesse with a sample size of 260 672 insurees. We used an anatomic-therapeutic-chemical classification (C08) to identify prescriptions for calcium-channel blockers. We determined from brand names and dosage forms whether nifedipine was prescribed in an immediate-release or sustained-release formulation. Among insurees over age 65, the prevalence of treatment with immediate-release and sustained-release nifedipine was 0.9% and 1.0%, respectively. Immediate-release nifedipine was usually (75%) given in a single administration. 46% of patients receiving immediate-release nifedipine also received another calcium-channel blocker. Patients who received immediate-release nifedipine tended to take more cardiovascular drugs than those who received sustained-release nifedipine (6 or more cardiovascular drugs were taken by 30% and 16%, respectively). Among all medical diagnoses related to hypertension, two were significantly more common among patients taking immediate-release nifedipine than among those taking sustained-release nifedipine: hypertensive crisis (OR 4.26, 95% CI 2.45-7.40) and hypertensive heart disease (OR 1.82, 95% CI 1.04-3.19). Our analysis demonstrates that immediate-release nifedipine is being prescribed to elderly patients in Germany, albeit mostly in a single administration. In view of the risks and the availability of alternative drugs, stricter adherence to the PRISCUS recommendations in this case should be stressed in continuing medical education.


Effects of conditioned media from murine lung cancer cells and human tumor cells on cultured myotubes.
Factors secreted from tumors/tumor cells are hypothesized to cause skeletal muscle wasting in cancer patients. We examined whether cancer cells secrete factors to promote atrophy by evaluating the effects of conditioned media (CM) from murine lung cancer cells and primary cultures of human lung tumor cells on cultured myotubes. We evaluated murine Lewis lung carcinoma (LLC) and KRASG12D cells, and primary cell lines derived from tumor biopsies from patients with lung cancer (hTCM; n = 6). In all experiments, serum content was matched across treatment groups. We hypothesized that CM from murine and human tumor cells would reduce myotube myosin content, decrease mitochondrial content, and increase mitochondrial reactive oxygen species (ROS) production. Treatment of myotubes differentiated for 7 days with CM from LLC and KRASG12D cells did not alter any of these variables. Effects of murine tumor cell CM were observed when myotubes differentiated for 4 days were treated with tumor cell CM and compared with undiluted differentiation media. However, these effects were not apparent if tumor cell CM treatments were compared with control cell CM or dilution controls. Finally, CM from human lung tumor primary cell lines did not modify myosin content or mitochondrial content or ROS production compared with either undiluted differentiated media, control cell CM, or dilution controls. Our results do not support the hypothesis that factors released from cultured lung cancer/tumor cells promote myotube wasting or mitochondrial abnormalities, but we cannot dismiss the possibility that these cells could secrete such factors in vivo within the native tumor microenvironment.


Properties of vestibular neurones projecting to neck segments of the cat spinal cord.
1. Vestibular neurones projecting to the upper cervical grey matter (vestibulocollic neurones) were identified by localized microstimulation in the C3 segment of the cat spinal cord.2. The neurones were found in the lateral (Deiters'), medial and descending nuclei bilaterally and projected to the spinal cord in the lateral and medial vestibulospinal tracts (LVST and MVST). Ipsilateral axons of Deiters' neurones were mostly in the LVST, axons of medial and descending neurones in the MVST; a few Deiters' neurones had axons in the MVST; some descending neurones had axons in the LVST. Most axons of contralateral neurones were in the MVST.3. The axons of 62% of ipsilateral vestibulocollic Deiters' neurones not only gave off a collateral to C3, but also extended as far as the cervical enlargement (;branching'); some of these neurones projected as far as the upper thoracic cord, almost none to the lumbar cord. Ipsilateral descending nucleus neurones branch in the same fashion, but there is no branching in the relatively small medial nucleus population.4. A large majority of vestibulocollic neurones receive monosynaptic excitation from the ipsilateral labyrinth and a number are inhibited by stimulation of the contralateral labyrinth (commissural inhibition). It is possible that commissural inhibition acts on a broad population of vestibular neurones involved in the control of eye, head and trunk movement.5. Vestibulocollic neurones do not make up a homogeneous population acting only on the neck. Instead it is likely that subpopulations, for example branching and non-branching neurones, have different functions.


Intrapopulation and temporal differences of phthalate concentrations in North Atlantic fin whales (Balaenoptera physalus).
The fin whale (Balaenoptera physalus) is a migratory filter-feeding species that is susceptible to ingest plastics while lunge feeding across the oceans. Plastic additives, such as phthalates, are compounds that are added to plastics to give them specific characteristics, such as flexibility. These so-called plasticizers are currently raising major concern because of their potential adverse effects on marine fauna. However, little is known about phthalate concentrations in tissues of baleen whales as well as their potential relation with biological variables (i.e., sex, body length and age) and their trends with time. In this study, we assessed the concentration of 13 phthalates in the muscle of 31 fin whales sampled in the feeding grounds off western Iceland between 1986 and 2015. We detected 5 of the 13 phthalates investigated, with di-n-butylphthalate (DBP), diethylphthalate (DEP) and bis(2-ethylhexyl) phthalate (DEHP) being the most abundant. None of the biological variables examined showed a statistically significant relationship with phthalate concentrations. Also, phthalate concentrations did not significantly vary over the 29-year period studied, a surprising result given the global scenario of increasing plastic pollution in the seas. The lack of time trends in phthalate concentration may be due in part to the fact that phthalates also originate from other sources. Although no adverse effects of phthalates on fin whales have been detected to date, further monitoring of these pollutants is required to identify potential toxic effects in the future.


Influence of scaffold stiffness on subchondral bone and subsequent cartilage regeneration in an ovine model of osteochondral defect healing.
In osteochondral defects, subchondral bone, as a load-bearing structure, is believed to be important for bone and cartilage regeneration. A stiff scaffold creates better conditions for bone formation and cartilage regeneration than does a softer one. Controlled laboratory study. Critical osteochondral defects were created in the femoral condyles of 24 sheep. Subchondral bone was reconstructed with a stiff scaffold or a modified softer one, with untreated defects serving as controls. The repair response was evaluated with mechanical, histological, and histomorphometrical techniques at 3 and 6 months postoperatively. The elastic modulus of regenerated fibrocartilage over the stiff scaffold tended to be higher than in the soft scaffold group (61% vs 46% of healthy cartilage) at 3 months. No difference was determined at 6 months; all were well below healthy cartilage. Treated defects showed substantial degradation of the soft scaffold with surrounding sclerotic bone at 3 and 6 months. In contrast, degradation of the stiff scaffold was slower and occurred together with continuous osseous replacement. Stiff scaffolds were found to improve bone regeneration. In contrast, soft scaffolds provided less support, and consequently subchondral bone became sclerotic. Although regenerated cartilage formed over the stiff scaffolds at 3 months, and these exhibited better mechanical properties than did the soft scaffold group, the mechanical properties in both treated groups were the same at 6 months, not dissimilar to that of tissue formed in the untreated specimens and inferior to native articular cartilage. The results imply that subchondral defect filling in clinical settings advances bone regeneration and should have a comparable stiffness to that of healthy subchondral bone rather than being too flexible. Degradation of resorbable materials and consequently the loss of stiffness may compromise the healing of critical defects.


Evaluating the Impact of Wildlife Shelter Management on the Genetic Diversity of Erinaceus europaeus and E. roumanicus in Their Contact Zone.
Hedgehogs are among the most abundant species to be found within wildlife shelters and after successful rehabilitation they are frequently translocated. The effects and potential impact of these translocations on gene flow within wild populations are largely unknown. In this study, different wild hedgehog populations were compared with artificially created "shelter populations", with regard to their genetic diversity, in order to establish basic data for future inferences on the genetic impact of hedgehog translocations. Observed populations are located within central Europe, including the species Erinaceus europaeus and E. roumanicus. Shelters were mainly hosting one species; in one case, both species were present syntopically. Apart from one exception, the results did not show a higher genetic diversity within shelter populations, indicating that individuals did not originate from a wider geographical area than individuals grouped into one of the wild populations. Two shelters from Innsbruck hosted individuals that belonged to two potential clusters, as indicated in a distance analysis. When such a structure stems from the effects of landscape elements like large rivers, the shelter management-related translocations might lead to homogenization across the dispersal barrier.


Nuclear mechanosensing: mechanism and consequences of a nuclear rupture.
The physical connections between the cytoskeletal system and the nucleus provide a route for the nucleus to sense the mechanical stress both inside and outside of the cell. Failure to withstand such stress leads to nuclear rupture, which is observed in human diseases. In this review, we will go through the recent findings and our current understandings of nuclear rupture. Starting with the triggers of nuclear rupture, including the aberrant nuclear lamina composition and the elevated actomyosin contractility. We will also discuss the role of ESCRT-III in nuclear rupture repair and the biological consequences of nuclear rupture, including the negative impacts on cellular compartmentalization, DNA damage, and cellular differentiation. Recent studies on nuclear rupture provide further insights into the direct mechanistic link between nuclear rupture and several pathological conditions. Such knowledge can guide us in developing potential therapeutic solutions for the patients.


The effects of coronary artery disease on the ventricular fibrillation threshold in man.
The ventricular fibrillation threshold (VFT) was measured in 28 patients at the time of cardiac surgery. The VFT was measured with a 100 Hz train of 24 rectangular pulses positioned across the ST segment and T wave. Current was applied to the epicardial surface of either ventricle with a bipolar electrode probe. In six patients, the normal right VFT was 24.3 +/- 5.2 mA, and in 10 patients the normal left VFT was 33.6 +/- mA (p less than 0.05). In 12 patients with greater than or equal to 75% obstruction of the left anterior descending coronary artery, the left VFT was 18.6 +/- 6.9 mA. This value was significantly less than the left VFT in patients without coronary artery disease (p less than 0.001). This study shows that the VFT can be measured in man and that coronary artery disease reduces this parameter.


A versatile low temperature solid-state synthesis of vanadium nitride (VN) via a "guanidinium-route": experimental and theoretical studies from the key-intermediate to the final product.
An easy and efficient method ("guanidinium route") to synthesize vanadium nitride (VN) is evaluated in this paper. Initially, ammonium m-vanadate was mixed with guanidinium carbonate, producing an important intermediate, ammonium m-vanadate (GmV), through a solid-state reaction. GmV was decomposed as a function of the temperature and studied with TGA, DRX, FT-Infrared, Temperature-Programmed Surface Reaction (TPSR) and DFT. We show that GmV is transformed into bulk crystalline VN below 800 °C. XPS, periodic DFT calculations, and elemental analyses show that the surface is not carbon-free.


Erratum: Value of Routine Sonographic Screening of Internal Jugular Vein to Detect Catheter Related Thrombosis in Intensive Care Unit.
[This corrects the article on p. 326 in vol. 23, PMID: 31406439.].


Plants 'cry' for help through acoustic signals.
Plants perceive sounds, while responses to these sounds were already known. A breakthrough is the discovery by Khait et al. that stressed plants emit various informative ultrasonic sound signals, which can be categorized according to plant species, stress type, and stress severity. This discovery may change how plants are cultivated.


Spirituality in palliative care: opportunity or burden?
The article questions an assumption in palliative care literature, namely that all patients have a spiritual dimension and that all staff can offer spiritual care. The article identifies spirituality as a particular kind of discourse. In late-modern Anglophone societies, this discourse arises from the experience of a particular generation and a particular segment of the population, namely those moving beyond formal religion; this segment is probably better represented among caring professionals than among dying patients. A four-fold typology of patients' approaches to religion/spirituality is developed, indicating the potential of differentiating between actual patients, rather than presuming a universal 'search for meaning'. This alternative approach may enhance opportunities for team working and reduce the likelihood of any one member of staff feeling spiritual care to be an unwelcome burden.


Evaluation of readmission ink as a marker for dispersal studies with the oriental fruit fly, Bactrocera dorsalis.
In this text we present a new marking dye, readmission ink, Blak-Ray, for the purpose of insect movement studies. The dye was tested in a controlled experiment with Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) in anticipation of a long distance movement study planned for the following year with the same species. 700 individuals of both sexes were marked with the dye and placed in holding containers. Both the percentage of mortality and the ease of dye detection were monitored throughout a five-week period. Results showed minimal fly mortality and exceptional ease of dye detection.


New metal-only Lewis pairs: elucidating the electronic influence of N-heterocyclic carbenes and phosphines on the dative Pt-Al bond.
The synthesis and full characterization of a new heteroleptic N-heterocyclic carbene (NHC)-phosphine platinum(0) complex and formation of its corresponding alane adduct is reported. The influence of the ligands on the Lewis basic properties was studied via multinuclear NMR-spectroscopy, X-ray analyses, and density functional theory (DFT) calculations. Consistently, the effect of changing the halogens upon the Lewis acid properties of aluminum halides was studied by X-ray analysis and DFT calculations.


Synthesis and structure of two novel trans-platinum complexes.
Here for the first time the synthesis and characterization of two new trans-platinum complexes, trans-[PtCl2{HN=C(OH)C6H5}2] (compound 1) and trans-[PtCl4(NH3){HN=C(OH)tBu}] (compound 2) [with tBu = C(CH3)3] are described. The structures have been characterized using nuclear magnetic resonance spectroscopy and X-ray single-crystal diffraction. In compound 1 the platinum cation, at the inversion center, is in the expected square-planar coordination geometry. It is coordinated to two chloride anions, trans to each other, and two nitrogen atoms from the benzamide ligands. The van der Waals interactions between the molecules produce extended two-dimensional layers that are linked into a three-dimensional structure through π...π intermolecular interactions. In compound 2 the platinum cation is octahedrally coordinated by four chloride anions and two nitrogen atoms from the pivalamide and ammine ligands, in trans configuration. The molecular packing is governed by intermolecular hydrogen bonds and van der Waals interactions.


Configurational coincidence among six phenomena: a comment on van Lier and Csathó (2006).
This study focuses on a configurational coincidence among six different illusions, two being motion illusions (reversed phi movement and phi movement), two being binocular stereo effects (Anstis-Rogers type and Gregory-Heard type), one being a position illusion (Gregory-Heard illusion), and one being an orientation illusion (Café-Wall-like tilt illusion). The stimuli of these six illusions or effects share the same configuration, in which a thin region is flanked by thick regions of different luminances. A phenomenological comparison is conducted with the use of dynamic demonstrations as well as static ones. In particular, the reversed phi movement is extensively discussed.


Role of Pituitary Adenylate-Cyclase Activating Polypeptide and Tac1 gene derived tachykinins in sensory, motor and vascular functions under normal and neuropathic conditions.
Pituitary Adenylate-Cyclase Activating Polypeptide (PACAP) and Tac1 gene-encoded tachykinins (substance P: SP, neurokinin A: NKA) are expressed in capsaicin-sensitive nerves, but their role in nociception, inflammation and vasoregulation is unclear. Therefore, we investigated the function of these neuropeptides and the NK1 tachykinin receptor (from Tacr1 gene) in the partial sciatic nerve ligation-induced traumatic mononeuropathy model using gene deficient (PACAP(-/-), Tac1(-/-), and Tacr1(-/-)) mice. Mechanonociceptive threshold of the paw was measured with dynamic plantar aesthesiometry, motor coordination with Rota-Rod and cutaneous microcirculation with laser Doppler imaging. Neurogenic vasodilation was evoked by mustard oil stimulating sensory nerves. In wildtype mice 30-40% mechanical hyperalgesia developed one week after nerve ligation, which was not altered in Tac1(-/-) and Tacr1(-/-) mice, but was absent in PACAP(-/-) animals. Motor coordination of the PACAP(-/-) and Tac1(-/-) groups was significantly worse both before and after nerve ligation compared to their wildtypes, but it did not change in Tacr1(-/-) mice. Basal postoperative microcirculation on the plantar skin of PACAP(-/-) mice did not differ from the wildtypes, but was significantly lower in Tac1(-/-) and Tacr1(-/-) ones. In contrast, mustard oil-induced neurogenic vasodilation was significantly smaller in PACAP(-/-) mice, but not in Tacr1(-/-) and Tac1(-/-) animals. Both PACAP and SP/NKA, but not NK1 receptors participate in normal motor coordination. Tachykinins maintain basal cutaneous microcirculation. PACAP is a crucial mediator of neuropathic mechanical hyperalgesia and neurogenic vasodilation. Therefore identifying its target and developing selective, potent antagonists, might open promising new perspectives for the treatment of neuropathic pain and vascular complications.


Domestic abuse in the Covid-19 pandemic: measures designed to overcome common limitations of trend measurement.
Research on pandemic domestic abuse trends has produced inconsistent findings reflecting differences in definitions, data and method. This study analyses 43,488 domestic abuse crimes recorded by a UK police force. Metrics and analytic approaches are tailored to address key methodological issues in three key ways. First, it was hypothesised that reporting rates changed during lockdown, so natural language processing was used to interrogate untapped free-text information in police records to develop a novel indicator of change in reporting. Second, it was hypothesised that abuse would change differentially for those cohabiting (due to physical proximity) compared to non-cohabitees, which was assessed via a proxy measure. Third, the analytic approaches used were change-point analysis and anomaly detection: these are more independent than regression analysis for present purposes in gauging the timing and duration of significant change. However, the main findings were largely contrary to expectation: (1) domestic abuse did not increase during the first national lockdown in early 2020 but increased across a prolonged post-lockdown period, (2) the post-lockdown increase did not reflect change in reporting by victims, and; (3) the proportion of abuse between cohabiting partners, at around 40 percent of the total, did not increase significantly during or after the lockdown. The implications of these unanticipated findings are discussed. The online version contains supplementary material available at 10.1186/s40163-023-00190-7.


Effect of glucocorticoid replacement therapy on bone mineral density in patients with Addison disease.
To study the influence of glucocorticoid replacement therapy on bone mineral density. Cross-sectional. University hospital in the Netherlands. 91 patients with Addison disease who had been receiving glucocorticoid replacement therapy for a mean of 10.6 years (range, 0.5 to 36.5 years). Bone mineral density of the lumbar spine and both femoral necks using a dual-energy x-ray absorptiometer and basal serum concentrations of adrenocorticotropin, gonadal hormones, and adrenal androgens. Decreased bone mineral density (less than 2 standard deviations [SD] of the mean value of an age-matched reference population) was found in 10 of 31 men (32%; 95% Cl, 17% to 51%) and in 4 of 60 women (7%; Cl, 2% to 16%). No statistically significant differences were found between men and women with regard to age, duration of glucocorticoid substitution, or glucocorticoid dose, either in absolute quantities or when expressed per kilogram of body weight. However, in men with decreased bone mineral density, the daily hydrocortisone dose per kilogram of body weight (0.43 +/- 0.08 mg/kg; mean +/- SD) was significantly (P = 0.032) higher than in men with normal bone mineral density (0.35 +/- 0.10 mg/kg). After correction for possible confounding variables, a significant linear correlation was found between hydrocortisone dose per kilogram of body weight and bone mineral density of the lumbar spine in the men (regression coefficient, -0.86; Cl, -1.60 to -0.13; P = 0.029) but not in the women. Long-term treatment with standard replacement doses of glucocorticoids may induce bone loss in men with Addison disease. Adjustment of glucocorticoid therapy to the lowest acceptable dose is mandatory in Addison disease, and regular measurement of bone mineral density may be helpful in identifying men at risk for the development of osteoporosis.


Patterns in forage fish mercury concentrations across Northeast US estuaries.
Biogeochemical conditions and landscape can have strong influences on mercury bioaccumulation in fish, but these effects across regional scales and between sites with and without point sources of contamination are not well understood. Normal means clustering, a type of unsupervised machine learning, was used to analyze relationships between forage fish (Fundulus heteroclitus and Menidia menidia) mercury (Hg) concentrations and sediment and water column Hg and methylmercury (MeHg) concentrations, ancillary variables, and land classifications within the sub-watershed. The analysis utilized data from 38 sites in 8 estuarine systems in the Northeast US, collected over five years. A large range of mercury concentrations and land use proportions were observed across sites. The cluster correlations indicated that for Fundulus, benthic and pelagic Hg and MeHg concentrations were most related to tissue concentrations, while Menidia Hg was most related to water column MeHg, reflecting differing feeding modes between the species. For both species, dissolved MeHg was most related to tissue concentrations, with sediment Hg concentrations influential at contaminated sites. The models considering only uncontaminated sites showed reduced influence of bulk sediment MeHg for both species, but Fundulus retained sediment drivers at some sites, with dissolved MeHg still highly correlated for both. Dissolved organic carbon (DOC), chlorophyll, land use, and other ancillary variables were of lesser importance in driving bioaccumulation, though DOC was strongly related within some clusters, likely in relation to dissolved Hg. Land use, though not of primary importance, showed relationships opposite to those observed in freshwater, with development positively correlated and forests and agriculture negatively correlated with tissue concentrations across clusters and species. Clusters were composed of sites from geographically distinct systems, indicating the greater importance of small scale drivers of MeHg formation and uptake into the food web over system or region-wide influences.


Seasonal variations in activity concentrations of 99Tc and 137Cs in the edible meat fraction of crabs and lobsters from the central Irish Sea.
Discharges of most radionuclides into the Irish Sea from the BNFL site at Sellafield have decreased over the past 20 years or so. For a few radionuclides, however, discharges have peaked more recently. Notably, operation of the Enhanced Actinide Removal Plant (EARP) since 1994 has led to an increase in discharges of (99)Tc, as a result of the treatment of previously stored waste, with consequent increases in (99)Tc activity concentrations in a number of marine species, particularly in crustaceans such as lobsters. Previous research has considered the significance of factors such as sex and body weight on radionuclide concentrations. The current project set out to investigate whether seasonal variations in radionuclide concentrations in crabs and lobsters occur, with particular emphasis on the dynamics of (99)Tc and (137)Cs. Organisms were obtained from a site off the Isle of Man, where radionuclide concentrations were measurable but the site was sufficiently distant from Sellafield that the radionuclides were well mixed in the water column and not likely to be influenced by the pulsed nature of discharges of (99)Tc. Crab and lobster samples were collected monthly, between February 2000 and February 2001. Fifteen or 16 individuals (evenly split as male and female) of each species were collected on each occasion. Seawater samples were also collected over the 12-month period. Activity concentrations of (99)Tc in the edible meat fraction (both brown and white meat) ranged from 0.23 to 2.46 Bq kg(-1) (fresh weight (fw)) in crabs and 124 to 216 Bq kg(-1) (fw) in lobsters, with no observed seasonal variations. Activity concentrations of (137)Cs in both crab and lobster were lower, ranging from <0.16 to 0.85 Bq kg(-1) for crab meat (fw) and <0.3 to 3.3 Bq kg(-1) for lobster meat (fw). A statistically significant increase in activity concentrations of (137)Cs in the meat was observed in the summer months for both crab and lobster. The cause has not been investigated but may be related to the laying down of energy reserves during the active feeding period over the summer. At all times, uptake of (99)Tc is higher in the brown meat fraction of both crabs and lobsters, whilst (137)Cs is more uniformly distributed. These results are used to discuss the implications for sampling and monitoring programmes.


Comparison of population genetic structures of common wild rice (Oryza rufipogon Griff.), as revealed by analyses of quantitative traits, allozymes, and RFLPs.
We investigated genetic diversity among and within natural populations of Asian common wild rice, Oryza rufipogon, from three different classes of data: quantitative traits, allozymes, and restriction fragment length polymorphisms (RFLPs). The seven populations examined showed polymorphism to varying degrees. The amount of intrapopulation variability appeared to be influenced not only by breeding system but also by the evolutionary history of each population. Interpopulation differentiation was clear, but different classes of data elucidated different patterns. Quantitative traits revealed ecotype differentiation into perennial and annual types of population, whereas allozyme and RFLP analyses revealed geographical differentiation among populations. These results suggest that the diversity patterns shown by quantitative trait analysis reflect mainly the occurrence of adaptive differentiation in response to habitat conditions and that those shown by allozyme and RFLP analyses reflect mainly the effect of isolation by distance. Population differentiation parameters (F(ST)) were highly variable among loci in allozymes as well as in RFLPs.


Graph theoretical analysis of magnetoencephalographic functional connectivity in Alzheimer's disease.
In this study we examined changes in the large-scale structure of resting-state brain networks in patients with Alzheimer's disease compared with non-demented controls, using concepts from graph theory. Magneto-encephalograms (MEG) were recorded in 18 Alzheimer's disease patients and 18 non-demented control subjects in a no-task, eyes-closed condition. For the main frequency bands, synchronization between all pairs of MEG channels was assessed using a phase lag index (PLI, a synchronization measure insensitive to volume conduction). PLI-weighted connectivity networks were calculated, and characterized by a mean clustering coefficient and path length. Alzheimer's disease patients showed a decrease of mean PLI in the lower alpha and beta band. In the lower alpha band, the clustering coefficient and path length were both decreased in Alzheimer's disease patients. Network changes in the lower alpha band were better explained by a 'Targeted Attack' model than by a 'Random Failure' model. Thus, Alzheimer's disease patients display a loss of resting-state functional connectivity in lower alpha and beta bands even when a measure insensitive to volume conduction effects is used. Moreover, the large-scale structure of lower alpha band functional networks in Alzheimer's disease is more random. The modelling results suggest that highly connected neural network 'hubs' may be especially at risk in Alzheimer's disease.


Structural and functional characteristics of irradiation damage to parotid glands in the miniature pig.
To evaluate the effects of a solitary megadose protocol of ionizing radiation (IR) on the structure and function of the miniature pig (minipig) parotid gland. Fourteen minipigs were subjected to either 15 or 20 Gy to one parotid gland with a linear accelerator, whereas another four minipigs served as non-IR controls. Salivary flow rates and salivary chemistries were measured pre-IR and 4 and 16 weeks post-IR. A quantitative assessment of gland weight and acinar area and detailed serum chemistry and hematologic analyses were also performed. Parotid flow rates decreased by approximately 50% either with 20 Gy at 4 weeks, or 15 Gy at 16 weeks post-IR. In the 20 Gy group, salivary flow rates were reduced by approximately 80% at 16 weeks post-IR. A significant decrease in salivary calcium and amylase and an increase of salivary potassium levels were found in both IR groups. There were also transient alterations in serum chemistry and hematology parameters post-IR. Parotid gland weights were significantly decreased (-50%) in the 15 and 20 Gy groups at 4 and 16 weeks post-IR. Additionally, the acinar cell area in glands of both IR groups was significantly reduced from that in control glands at both the 4 and 16 weeks time points. Structural changes in salivary gland parenchyma occurred relatively early after IR, whereas the alterations in salivary output were relatively delayed. Further, reductions in salivary flow were not proportional to acinar cell area loss. Together, these findings suggest that nonparenchymal IR damage likely contributes to IR-induced salivary hypofunction.


Actin dynamics is essential for myosin-based transport of membrane organelles.
Actin filaments that serve as "rails" for the myosin-based transport of membrane organelles [1-4] continuously turn over by concurrent growth and shortening at the opposite ends [5]. Although it is known that dynamics of actin filaments is essential for many of the actin cytoskeleton functions, the role of such dynamics in myosin-mediated organelle transport was never studied before. Here, we addressed the role of turnover of actin filaments in the myosin-based transport of membrane organelles by treating cells with the drugs that suppress actin-filament dynamics and found that such a suppression significantly inhibited organelle transport along the actin filaments without inhibiting their intracellular distribution or the activity of the myosin motors. We conclude that dynamics of actin filaments is essential for myosin-based transport of membrane organelles and suggest a previously unknown role of actin-filament dynamics in providing the "rails" for continuous organelle movement resulting in the increased distances traveled by membrane organelles along the actin filaments.


Metabolic alterations in K562 cells exposed to taxol and tyrphostin AG957: 1H NMR and biochemical studies.
K562 cells exposed for 3 h to taxol or taxol plus tyrphostin AG957 exhibited a significant variation in the concentration of the water-soluble metabolites glutathione, myo-inositol and phosphorylcholine, as evaluated by (1)H NMR up to 72 h incubation in drug-free medium. Cells treated with both drugs showed an increase of glutathione and glutathione reductase at 24 h and a sharp decrease of myo-inositol between 8 and 24 h. Phosphorylcholine increased at 8 h both in taxol and taxol plus AG957-treated cells, which was then abruptly inverted to a significantly lower concentration at 24 h, subsequently increasing again to values higher than those found in taxol-treated and control cells. All the above reported effects were lacking in cells exposed to AG957 alone. These modifications, despite the enhancement of the overall apoptotic cascade in taxol plus AG957-treated cells, can be related to the activation of cellular detoxification mechanisms, to the correct osmolarity maintenance, and to alterations of phospholipid metabolism.


Inorganic phosphate regulates multiple genes during osteoblast differentiation, including Nrf2.
The process of osteoblast differentiation and matrix mineralization requires a rise in alkaline phosphatase enzymatic activity resulting in the generation of free phosphate. The ability of inorganic phosphate to regulate gene transcription and cellular function represents a potentially novel extracellular signaling mechanism. Using microarray analysis we have identified a discrete set of genes that are either positively or negatively regulated by increased phosphate in MC3T3-E1 cells. The genes downregulated by phosphate encode for osteoblast-related extracellular factors such as collagens, periostin, and decorin. The genes increased by phosphate encode a novel group of transcription factors that may be important in the later stages of osteoblast development in which the environment is high in phosphate. The transcription factor Nrf2 is one such gene. Elevated phosphate levels stimulate an increase in Nrf2 RNA that is not blocked by the translation inhibitor cycloheximide, suggesting that Nrf2 is an immediate response gene. Cloning of the murine nrf2 promoter reveals that elevated phosphate produces an increase in promoter activity that is both time and dose dependent. This analysis reveals multiple genes regulated by the increase in phosphate associated with osteoblast differentiation, adding to our understanding of the intricate communication between osteoblasts and their extracellular environment.


Involvement of the mitochondrial calcium uniporter in cardioprotection by ischemic preconditioning.
The objective of the present study was to determine whether the mitochondrial calcium uniporter plays a role in the cardioprotection induced by ischemic preconditioning (IPC). Isolated rat hearts were subjected to 30 min of regional ischemia by ligation of the left anterior descending artery followed by 120 min of reperfusion. IPC was achieved by two 5-min periods of global ischemia separated by 5 min of reperfusion. IPC reduced the infarct size and lactate dehydrogenase release in coronary effluent, which was associated with improved recovery of left ventricular contractility. Treatment with ruthenium red (RR, 5 microM), an inhibitor of the uniporter, or with Ru360 (10 microM), a highly specific uniporter inhibitor, provided cardioprotective effects like those of IPC. The cardioprotection induced by IPC was abolished by spermine (20 microM), an activator of the uniporter. Cyclosporin A (CsA, 0.2 microM), an inhibitor of the mitochondrial permeability transition pore, reversed the effects caused by spermine. In mitochondria isolated from untreated hearts, both Ru360 (10 microM) and RR (1 microM) decreased pore opening, while spermine (20 microM) increased pore opening which was blocked by CsA (0.2 microM). In mitochondria from preconditioned hearts, the opening of the pore was inhibited, but this inhibition did not occur in the mitochondria from hearts treated with IPC plus spermine. These results indicate that the mitochondrial calcium uniporter is involved in the cardioprotection conferred by ischemic preconditioning.


ANA (+) ANCA (+) systemic vasculitis associated with the use of minocycline: case-based review.
Minocycline is a synthetic tetracycline-derived antibiotic with significant anti-inflammatory properties that may benefit patients with rheumatoid arthritis. Surprisingly, chronic exposure to minocycline can also cause a breach in immunologic tolerance resulting in a variety of autoimmune syndromes such as drug-induced lupus or autoimmune hepatitis. Vasculitis, most commonly resembling cutaneous polyarteritis nodosa, has also been seen in patients taking this drug. Herein, we present a case of biopsy-proven systemic vasculitis presenting as an ANA (+) ANCA (+) polyarteritis nodosa-like syndrome in a male patient who was taking minocycline for his acne for approximately 2 years. Patient initially presented with constitutional symptoms such as profound weight loss and fatigue, along with myalgias, oligoarticular arthritis, and livedo reticularis. About 2 months later, he developed a severe left testicular pain. Biopsy showed vasculitis complicated with the infarction of the left testis. Angiography revealed microaneurysms in the renal and splenic circulation. Stopping the offending drug, along with the short course of prednisone and hydroxychloroquine, resulted in prompt resolution of his symptoms. We additionally present a comprehensive review of biopsy-proven cases of vasculitis associated with chronic minocycline treatment focusing on its pathogenesis and clinical manifestations.


Capsaicin modulates proliferation, migration, and activation of hepatic stellate cells.
Capsaicin, the active component of chili pepper, has been reported to have antiproliferative and anti-inflammatory effects on a variety of cell lines. In the current study, we aimed to investigate the effects of capsaicin during HSC activation and maintenance. Activated and freshly isolated HSCs were treated with capsaicin. Proliferation was measured by incorporation of EdU. Cell cycle arrest and apoptosis were investigated using flow cytometry. The migratory response to chemotactic stimuli was evaluated by a modified Boyden chamber assay. Activation markers and inflammatory cytokines were determined by qPCR, immunocytochemistry, and flow cytometry. Our results show that capsaicin reduces HSC proliferation, migration, and expression of profibrogenic markers of activated and primary mouse HSCs. In conclusion, the present study shows that capsaicin modulates proliferation, migration, and activation of HSC in vitro.


Identification of heme macrocycle type by near-infrared magnetic circular dichroism spectroscopy at cryogenic temperatures.
The electron paramagnetic resonance (EPR) and near-infrared magnetic circular dichroism (MCD) spectra of the azide and cyanide adducts of nitrimyoglobin and hydroperoxidase II from Escherichia coli have been measured at cryogenic temperatures. For the first time, ligand-to-metal charge-transfer transitions in the near-infrared have been observed for an Fe(III)-chlorine system. It is shown that near-ultraviolet-to-visible region electronic spectra of 'green' hemes such as these are an unreliable indicator of macrocycle type. However, the combined application of EPR and near-infrared MCD spectroscopies clearly distinguishes between the porphyrin-containing nitrimyoglobin and the chlorine-containing hydroperoxidase II.


[Taxonomic composition and landscape-biotopical distribution of black flies (Diptera, Simuliidae) in the Khemchik river basin (Western Tuva)].
Taxonomoc composition and altitude distribution of black flies have been investigated in the top and middle stream of the Khemchik river, Western Tuva. Similarity with adjacent territories by the species composition is revealed. A dependence of the blackfly abundance on the stream characteristics (water temperature, flow velocity, grading of soil, electric conductivity of water, and presence of epibioses), as well as on the hydrobiology characteristics of the localities is shown. The comparison of blackfly complexes from the upper part of the Sayano-Shushenskoe reservoir and the Todzha depression, Tuva, is carried out.


Limitations in randomised controlled trials evaluating drug effects in mania.
Considering the increasing number of drugs evaluated for mania in randomised controlled trials (RCTs) and the potential discrepancies between recommendations based on RCTs and the antimanic treatment given in clinical practice, this paper addresses some issues related to RCTs on drug effects in mania. One major question raised in the paper is to what extent selection prior to the point of randomisation in RCTs in mania may limit the applicability of study results to patients seen in ordinary clinical practice. Although such limitations in generalisability can be difficult to investigate empirically, it is emphasised that they should be openly discussed in the reports of RCTs. Another major focus is the issue of evaluation and interpretation of outcome, including a discussion of various response criteria based on mania rating scale scores. It is pointed out that essential criteria of dimensionality have only been sufficiently evaluated for the Bech-Rafaelsen Mania Rating Scale, although the fulfilment of such criteria are prerequisites for adding up the item scores to a total score reflecting the severity of mania. It is suggested that response defined as a decline in mania score below a certain limit may have some advantages over the commonly used 50% reduction criterion. The issues arising from the unusual high drop-out rates of around 50% are also addressed. Despite the fact that we need rigorous placebo-controlled trials to establish antimanic efficacy of new compounds, we also need large scale pragmatic studies using broad inclusion criteria, comparing the various treatments, alone or in combination, to investigate how they work in clinical practice. These studies maybe randomised but open and use simple but relevant outcome measures.


Ablation of RIP3 protects from dopaminergic neurodegeneration in experimental Parkinson's disease.
Parkinson's disease (PD) is driven by dopaminergic neurodegeneration in the substantia nigra pars compacta (SN) and striatum. Although apoptosis is considered the main neurodegenerative mechanism, other cell death pathways may be involved. In this regard, necroptosis is a regulated form of cell death dependent on receptor interacting protein 3 (RIP3), a protein also implicated in apoptosis and inflammation independently of its pro-necroptotic activity. Here, we explored the role of RIP3 genetic deletion in in vivo and in vitro PD models. Firstly, wild-type (Wt) and RIP3 knockout (RIP3ko) mice were injected intraperitoneally with MPTP (40 mg/kg, i.p.), and sacrificed after either 6 or 30 days. RIP3ko protected from dopaminergic neurodegeneration in the SN of MPTP-injected mice, but this effect was independent of necroptosis. In keeping with this, necrostatin-1s (10 mg/kg/day, i.p.) did not afford full neuroprotection. Moreover, MPTP led to DNA fragmentation, caspase-3 activation, lipid peroxidation and BAX expression in Wt mice, in the absence of caspase-8 cleavage, suggesting intrinsic apoptosis. This was mimicked in primary cortical neuronal cultures exposed to the active MPTP metabolite. RIP3 deficiency in cultured cells and in mouse brain abrogated all phenotypes. Curiously, astrogliosis was increased in the striatum of MPTP-injected Wt mice and further exacerbated in RIP3ko mice. This was accompanied by absence of microgliosis and reposition of glial cell line-derived neurotrophic factor (GDNF) levels in the striata of MPTP-injected RIP3ko mice when compared to MPTP-injected Wt mice, which in turn showed a massive GDNF decrease. RIP3ko primary mixed glial cultures also presented decreased expression of inflammation-related genes upon inflammatory stimulation. These findings hint at possible undescribed non-necroptotic roles for RIP3 in inflammation and MPTP-driven cell death, which can contribute to PD progression.


Phakomatosis pigmentovascularis with Sturge-Weber syndrome: a case report.
We describe here a Turkish girl with pigmentovascularis type 2b, consisting of disseminated Mongolian-spot-like maculae and unilateral Sturge-Weber angiomatosis.


Liver metastases, a rare cause of portal hypertension and stoma bleeding. Brief review of literature.
Portal hypertension is an unusual complication of liver metastases, which is frequently occurring in malignant disease. Portal hypertension may cause oesophageal varices and also stoma varices (colostomy and ileostomy). Oesophageal varices and bleeding from these varices have been frequently reported in literature. Stomal varices have also been reported in literature mostly associated with liver cirrhosis. These stomal varices lead to the massive bleeding causing morbidity and mortality. Portal hypertension is a pathological increase in portal pressure gradient (the difference between pressure in the portal and inferior vena cava veins). It is either due to an increase in portal blood flow or an increase in vascular resistance or combination of both. In liver cirrhosis, the primary factor leading to portal hypertension is increase in portal blood flow resistance and later on development of increased portal blood flow. It has been postulated that in liver metastasis the increase in portal flow resistance occurs at any site within portal venous system as a consequence of mechanical architectural disturbance. We report a case of a 64 year old gentleman who developed portal hypertension due to secondary metastases from colorectal cancer. He subsequently developed bleeding varices in his end colostomy. We believe that the combination of extensive metastases and chemotherapy induced portal hypertension in our patient. Our case and other literature review highlight that the recurrent bleeding stoma associated with colorectal cancer should be investigated for portal hypertension.


Pathogenicity of Mycoplasma capricolum in sheep after experimental infection.
Mycoplasma (M.) capricolum has been frequently isolated from diseased as well as from healthy sheep in Morocco. In order to determine its pathogenicity for sheep, experiments were performed in three trials with the Moroccan isolate 012. The following results were obtained: Ewes inoculated intramammarily developed acute mastitis; the organism was transmitted to lambs suckling these ewes. Only a mild mastitis appeared after a second inoculum, performed 5 weeks after the first. Young lambs, 1 month of age, fed four times with M. capricolum broth culture, died during the septicemic phase, showing a generalized septicemia, polyarthritis and a diffuse interstitial pneumonia. Lambs, 2 and 3 months of age inoculated intrabronchially developed a disease, which appeared to be age and weight dependent. The more susceptible animals died within 1-2 weeks after infection. The older lambs recovered gradually from the disease, which was characterized by pneumonia, conjunctivitis and arthritis. A significant increase of antibodies against M. capricolum developed in the older animals in the complement fixation test.


Decreased bone mineral density with off-label use of tenofovir in children and adolescents infected with human immunodeficiency virus.
5 of 6 children infected with human immunodeficiency virus (HIV) receiving Tenofovir disoproxil fumarate (TDF) experienced absolute decreases in bone mineral density (BMD). 2 pre-pubertal subjects experienced >6% BMD decreases. 1 subject was the smallest child and experienced a 27% decrease, necessitating withdrawal of TDF. Subsequently, her BMD recovered. Monitoring of children infected with HIV who require treatment with TDF is warranted.


Use of bronchopulmonary lavage for eliminating inhaled fume particles from a patient with arc welder's lung.
A 42-year-old man, who had worked as a welder for 20 years, was admitted to our hospital complaining of a dry cough. A chest radiograph showed diffuse small nodular shadows and chest computed tomography revealed small patchy opacities. A transbronchial lung biopsy specimen showed welding fume particles mainly located in alveolar space with mild fibrosis of alveolar septa. In order to prevent further fibrosis, bronchopulmonary lavage (BPL) was performed to eliminate the fume particles. The amount of iron particles derived from the total lavage fluid was 911.7 mg.


Sex differences in schizophrenia: a longitudinal methylome analysis.
DNA methylation analysis at the genome-wide level is a useful tool to explore potential sex differences in SCZ patients. The primary aim of the current study was to identify differentially methylated regions of DNA between males and females with schizophrenia. We collected DNA samples from 134 schizophrenia patients to measure genome-wide methylation at single-base resolution in 96 males and 38 females. We further repeated the analysis in 13 subjects (9 females, 4 males) to confirm the sex differences and to reduce the effect of potential confounders. The longitudinal methylation analysis found significant replication of several genes across the genome. These genes included RFTN1, TLE1, DAZL, PRR4, UTP14C, RNU12, and LOC644649. The overall results showed robust association between autosomal CpG sites and sex. Longitudinal methylation analysis can be used as internal replication to confirm epigenetic variants that are stable over time.


Downbeat nystagmus in two siblings with spinocerebellar ataxia type 6 (SCA 6).
We report two siblings with spinocerebellar ataxia type 6 (SCA 6), both showing downbeat nystagmus (DBN) as a predominant clinical feature. Familial hemiplegic migraine (FHM), episodic ataxia type 2 (EA-2) and SCA 6 are allelic disorders, and interestingly, the occasional presence of DBN in EA-2 was reported. Our observations suggest that common molecular mechanisms might underlie DBN in FHM, EA-2 and SCA 6. Then, these disorders should be kept in mind in diagnosing patients with DBN.


Eucalyptus pollen allergy and asthma in children: a cross-sectional study in South-East Queensland, Australia.
To investigate Eucalyptus (gum tree) pollen allergy in children in relation to geography, particularly vegetation, and its relationship to asthma. Males (n = 180) and females (n = 200) aged 9 to 14 participated. Some were healthy (asymptomatic), some had asthma, and some had other symptoms associated with atopy. School students were from three urban coastal schools and one school from a nearby semi-rural elevated area (range) near Brisbane, Australia. Coastal and range locations featured different distributions of Myrtaceae family vegetation (including Eucalyptus, Melaleuca, Leptospermum species). Skin prick test (SPT) responses to 15 commercial allergens were compared. As well, responses from coast versus range groups, and 'asthma' (n = 97) versus 'healthy' status (n = 107) groups, were compared. SPT responses (≥3mm wheal diameter) indicate that children with asthma are 31.1 times more likely to be allergic to Eucalyptus pollen extract (OR: 31.1; 95%CI 4.1- 235.7) compared to healthy children. Dust mite (p = .018), Eucalyptus (p = .046) and cockroach (p = .047) allergen SPT responses (wheals ≥3mm) were significantly greater in participants located on the coast versus range as determined by Fisher's Exact Test (α .05). For each location, percentage of positive responses (wheals ≥3mm) was greatest for 'dust mite' (30.9%-46%), 'cockroach' (18.1% -35%) and 'Bermuda grass' (10.6%-19.4%). The results support the hypothesis that proximity to Myrtaceae vegetation is related to positive SPT response and that Eucalyptus is an important allergen for children with asthma. Substantial response to olive allergen, in the absence of olive trees, suggests that the response may be driven by substances in other plants, perhaps Melaleuca quinquenervia, which abounds in coastal areas. Response to Eucalyptus allergen indicates that changes in gardening practice in schools and public areas may be appropriate. The findings pose validity questions regarding the use of some commercial allergens due to cross-reactive responses and the sources of those responses.


Should routine risk reduction procedures for the prevention and control of pandemics become a standard in all oncological outpatient clinics? The prospective COVID-19 cohort study: protect-CoV.
Limited knowledge exists on the effectiveness of preventive preparedness plans for the care of outpatient cancer patients during epidemics or pandemics. To ensure adequate, timely and continuous clinical care for this highly vulnerable population, we propose the establishment of preventive standard safety protocols providing effective early phase identification of outbreaks at outpatient cancer facilities and communicating adapted standards of care. The prospective cohort study Protect-CoV conducted at the LMU Klinikum from mid-March to June 2020 investigated the effectiveness of a rapid, proactive and methodical response to protect patients and interrupt SARS-CoV-2 transmission chains during the first pandemic wave. The implemented measures reduced the risk of infection of individual cancer patients and ensured safe adjunctive infusion therapy in an outpatient setting during the early COVID-19 pandemic. In addition to the immediate implementation of standard hygiene procedures, our results underscore the importance of routine PCR testing for the identification of asymptomatic or pre-symptomatic COVID-19 cases and immediate tracing of positive cases and their contacts. While more prospective controlled studies are needed to confirm these results, our study illustrates the importance of including preventative testing and tracing measures in the standard risk reduction procedures at all out patient cancer centers.


[Impact of age on the clinical response of patients with CML treated with imatinib].
Treatment of chronic myeloid leukemia (CML) with tyrosine kinase inhibitors (TKI) has significantly changed the survival of patients of all age categories. In our work, we have tried retrospectively to analyze a therapeutic response and toxicity of imatinib according to age of patients ( 60 years vs > 60 years). Patients have started treatment with imatinib in 2001-2013 in our department. The duration of treatment was at least 18 months. The whole group contains 103 patients (52 women, 51 men) with a median age at diagnosis of CML 55 years (range 19-88 years). The first group includes 68 the younger patients (31 women, 37 men) with a median age at diagnosis 43 years (range 19-59 years) and the second group 35 the elderly patients (21 women, 14 men) with a median age at diagnosis 70 years (range 61-86 years). Median imatinib dose in both groups was the same - 400 mg per day (range 200-600 mg). In the elderly patients we observed higher rates of hematological toxicities (p = 0.0059). After 12 months treatment, a complete cytogenetic response (CCyR) was achieved in 90 % of younger patients and in 74 % of the elderly patients, respectively. Major molecular response (MMR) at 18 months treatment was achieved in 79 % of the younger group and in 63 % of the elderly group. The results represent in both groups very good therapeutic response independent of age of patients at the time of CML diagnosis.


Effects of growth hormone on the function of beta-adrenoceptor subtypes in rat adipocytes.
The influence of growth hormone (GH) on the regulation of lipolytic response to specific agonists to beta-adrenoceptors and several post-receptor steps in the lipolytic cascade were investigated. Adipose tissues from rats were incubated with or without GH (1.38 nM). After a 24-hour incubation, isolated adipocytes were prepared for different assays. Rats were hypophysectomized. One week after operation, L-thyroxine and hydrocortisone acetate was given to hypophysectomized rats. One group of rats was treated with GH (1.33 mg/kg, daily). After 1 week of hormonal treatment, adipose tissues were removed for different studies. GH treatment increased both basal lipolysis and lipolytic sensitivity to dobutamine and CGP 12177 in adipocytes. The lipolytic sensitivity to terbutaline was not influenced by GH treatment. GH treatment increased the maximal lipolytic response to dobutamine and CGP 12177, but not to terbutaline as determined with absolute values of lipolysis. Forskolin-induced lipolysis was increased by addition of GH to tissues. Moreover, GH treatment resulted in enhanced expression of hormone-sensitive lipase. GH treatment in hypophysectomized rats influenced neither the expressions of G alpha s protein and cholera toxin-catalyzed adenosine diphosphate-ribosylation of G alpha s protein, nor cholera toxin-induced 3',5'-cyclic adenosine monophosphate accumulation. However, the expression of G alpha i protein was decreased after GH treatment. These and previous results suggest that GH increases lipolysis in rat adipocytes partly through the beta-adrenergic system, including increases in both beta(1)- and beta(3)-adrenergic receptor function, and partly through enhanced adenylate cyclase function, and expression of hormone-sensitive lipase, perhaps via a decrease in G alpha i protein expression.


Non-invasive measurement of baroreflex during Valsalva maneuver: Evaluation of alpha and beta-adrenergic components.
To evaluate alpha and beta components of adrenergic baroreflex sensitivity (BRSa) in Valsalva maneuver (VM). BRSa was studied in 89 healthy subjects aged 30±13 [16-75] years. Subjects were divided into three groups per blood pressure (BP) patterns associated with relatively balanced or increased alpha-adrenergic modulation: (1) BAR (n=43) - Balanced Autonomic Response with a BP dip below baseline in late phase II (IIL) and recovery in phase IV; (2) SAR (n=16) - Suppressed Autonomic Response with a non-dipping BP; and (3) AAR (n=30) - Augmented Autonomic Response with a BP recovery in phase IIL. Discrete (alpha and beta) BRSa formulae were produced using alpha- or beta-adrenergic phases: α-BRSa (phase IIL) and β-BRSa (phase IV), respectively. Discrete BRSa were studied to determine potential correlations to BRSa1 (validated BRSa evaluation) and evaluated for reliability. Patterns with higher α-adrenergic influence showed correlation between α-BRSa and BRSa1 (AAR: r=0.447, p<0.05; SAR: r=0.774, p<0.01). BAR showed correlation between β-BRSa and BRSa1 (r=-0.566, p<0.01), and α- and β-adrenergic coefficients (r=-0.381, p<0.05). Discrete BRSa were more reliable than BRSa1 (n=33; p<0.05). Discrete BRSa are reproducible and correlated with BRSa1. If validated, discrete BRSa may differentiate physiologic variances and vague dysautonomia.


The experience of faculty practice: developing and evaluating a newly established program.
While the notion of faculty practice, that is clinical practice by an academic in a health service environment, is not new, Australian Universities have been slow in providing practice environments where academics' theoretical understanding is informed through the service environment. Although there is a plethora of published academic opinion on the benefits, there is a dearth of meaningful data describing the subjective experience of academics that participate in faculty practice. Developing an understanding of the issues academics experience, while on faculty practice, provides a vital opportunity for those seeking to adopt a faculty practice model in their institution. The paper describes the genesis of the faculty practice program and outlines both the benefits and challenges that were encountered during implementation. A program evaluation conducted by an independent consultant indicated that all faculty practice participants found the process to be empowering and revitalising, despite their initial apprehension. The personal and professional gains achieved while on faculty practice were considered to compensate for the additional workload involved. The immediate dividends of enhanced self-esteem, classroom practices and credibility with students for faculty practice participants were outcomes achieved.


CNTF-mediated preactivation of astrocytes attenuates neuronal damage and epileptiform activity in experimental epilepsy.
Activated astrocytes display a broad spectrum of properties, ranging from neuroprotection to active contribution to demise of neural tissue. To investigate if activation of astrocytes by a single, defined stimulus enhances neuroprotective properties, we tested whether injection of ciliary neurotrophic factor (CNTF) can ameliorate epilepsy-related brain damage. Intrahippocampal CNTF injection in mice induced a rapid (within 2 days) and persistent (3 weeks) activation of astrocytes reflected by strong upregulation of glial fibrillary acidic protein (GFAP) mRNA synthesis and GFAP immunoreactivity. Moreover, CNTF signaling via phosphorylation and nuclear translocation of STAT3 (signal transducer and activator of transcription 3) was specifically activated in GFAP-positive astrocytes. CNTF-mediated activation of astrocytes 2 days prior to an epileptogenic intrahippocampal injection of kainate (KA) resulted in strongly reduced cell death in the hilus and CA3 region of the hippocampus, revealed by Fluoro-Jade B staining. Granule cell dispersion, the pathological widening of the granule cell layer, was also significantly reduced 16 days after KA injection. Importantly, intrahippocampal in vivo recordings 3 weeks after KA injection showed that the occurrence of high frequency oscillations (fast ripples, FR), a surrogate marker for epileptic activity, was significantly reduced in CNTF+KA-injected mice as compared to KA-injected animals. However, when CNTF was applied in the chronic epileptic phase at 3 weeks after KA injection, no reduction of FR activity was observed. In summary, our results indicate that the activation of astrocytes prior to an excitotoxic injury effectively reduces neuronal damage and the severity of epileptiform activity, whereas activation in the chronic phase is no longer protective.


Maternal and neonatal circulating markers of metabolic and cardiovascular risk in the metformin in gestational diabetes (MiG) trial: responses to maternal metformin versus insulin treatment.
This study was designed to compare glucose, lipids, and C-reactive protein (CRP) in women with gestational diabetes mellitus treated with metformin or insulin and in cord plasma of their offspring and to examine how these markers relate to infant size at birth. Women with gestational diabetes mellitus were randomly assigned to metformin or insulin in the Metformin in Gestational Diabetes trial. Fasting maternal plasma glucose, lipids, and CRP were measured at randomization, 36 weeks' gestation, and 6-8 weeks postpartum as well as in cord plasma. Women with available cord blood samples (metformin n = 236, insulin n = 242) were included. Maternal plasma triglycerides increased more from randomization to 36 weeks' gestation in women treated with metformin (21.93%) versus insulin (9.69%, P < 0.001). Maternal and cord plasma lipids, CRP, and neonatal anthropometry did not differ between treatments. In logistic regression analyses adjusted for confounders, the strongest associations with birth weight >90th centile were maternal triglycerides and measures of glucose control at 36 weeks. There were few differences in circulating maternal and neonatal markers of metabolic status and no differences in measures of anthropometry between the offspring of women treated with metformin and the offspring of women treated with insulin. There may be subtle effects of metformin on maternal lipid function, but the findings suggest that treating gestational diabetes mellitus with metformin does not adversely affect lipids or CRP in cord plasma or neonatal anthropometric measures.


Crystal structure of PvuII endonuclease reveals extensive structural homologies to EcoRV.
The crystal structure of the dimeric PvuII restriction endonuclease (R.PvuII) has been determined at a resolution of 2.4A. The protein has a mixed alpha/beta architecture and consists of two subdomains. Despite a lack of sequence homology, extensive structural similarities exist between one R.PvuII subdomain and the DNA-binding subdomain of EcoRV endonuclease (R.EcoRV); the dimerization subdomains are unrelated. Within the similar domains, flexible segments of R.PvuII are topologically equivalent to the DNA-binding turns of R.EcoRV; potential catalytic residues can be deduced from the structural similarities to R.EcoRV. Conformational flexibility is important for the interaction with DNA. A possible classification of endonuclease structures on the basis of the positions of the scissile phosphates is discussed.


Evolution of the epidemiological surveillance indicators of the main sexually transmitted infections in Spain: A retrospective observational study (2011-2019).
One of the most relevant aspects of sexual health promotion is the need to carry out health education by promoting a healthy affective-sexual education, with the capacity and resources to prevent sexually transmitted infections (STIs) and unwanted pregnancies. The aim of this study was to collect the evolution of epidemiological surveillance indicators of the main STIs, stratified by autonomous community after the publication of the Sexual and Reproductive Health Strategy in Spain in 2011 and until 2019. A retrospective observational study of the Spanish population with STIs from 2011-2019. The study variables were indicators of the main STIs, including annual rates of notification of infections per 100,000 inhabitants. There is a lack of homogeneous and systematic epidemiological surveillance records for STIs. Overall, in Spain there was an upward trend in STIs, except for HIV, which showed a clear decrease. Despite existing regulations, strategies, and sex education programs in Spain, the main STIs have been increasing over the years, and Spain is one of the countries with the highest rates in Europe.


Selection of housekeeping genes for gene expression studies in larvae from flatfish using real-time PCR.
Flatfish metamorphosis involves major physiological and morphological changes. Due to its importance in aquaculture and as a model for developmental studies, some gene expression studies have focused on the understanding of this process using quantitative real-time PCR (qRT-PCR) technique. Therefore, adequate reference genes for accurate normalization are required. The stability of 12 potential reference genes was examined during larval development in Senegalese sole (Solea senegalensis) and Atlantic halibut (Hippoglossus hippoglossus) to determine the most suitable genes for qRT-PCR analysis. Transcription levels of genes encoding beta-Actin (ACTB), glyceraldehyde-3P-dehydrogenase (GAPDH), annexin A2 (ANXA2), glutathione S-transferase (GST), ornithine decarboxylase (ODC), hypoxanthine phosphoribosyltransferase (HPRT1), ubiquitin (UBQ), elongation factor 1 alpha (eEF1A1), 18S ribosomal RNA, and the ribosomal proteins S4 (RPS4) and L13a (RPL13a) were quantitated. Two paralogous genes for ACTB were analyzed in each of both flatfish species. In addition, two paralogous genes for GAPDH were studied in Senegalese sole. RPL13a represented non-orthologous genes between both flatfish species. GeNorm and NormFinder analyses for expression stability revealed RPS4, UBQ and eEF1A1 as the most stable genes in Senegalese sole, Atlantic halibut and in a combined analysis. In all cases, paralogous genes exhibited differences in expression stability. This work suggests RPS4, UBQ, and eEF1A1 genes as useful reference genes for accurate normalization in qRT-PCR studies in Senegalese sole and Atlantic halibut larvae. The congruent results between both species in spite of the drastic differences in larval development suggest that selected housekeeping genes (HKGs) could be useful in other flatfish species. However, the finding of paralogous gene copies differentially expressed during development in some HKGs underscores the necessity to identify orthologous genes.


Identification of CASZ1 NES reveals potential mechanisms for loss of CASZ1 tumor suppressor activity in neuroblastoma.
As a transcription factor, localization to the nucleus and the recruitment of cofactors to regulate gene transcription is essential. Nuclear localization and nucleosome remodeling and histone deacetylase (NuRD) complex binding are required for the zinc-finger transcription factor CASZ1 to function as a neuroblastoma (NB) tumor suppressor. However, the critical amino acids (AAs) that are required for CASZ1 interaction with NuRD complex and the regulation of CASZ1 subcellular localization have not been characterized. Through alanine scanning, immunofluorescence cell staining and co-immunoprecipitation, we define a critical region at the CASZ1 N terminus (AAs 23-40) that mediates the CASZ1b nuclear localization and NuRD interaction. Furthermore, we identified a nuclear export signal (NES) at the N terminus (AAs 176-192) that contributes to CASZ1 nuclear-cytoplasmic shuttling in a chromosomal maintenance 1-dependent manner. An analysis of CASZ1 protein expression in a primary NB tissue microarray shows that high nuclear CASZ1 staining is detected in tumor samples from NB patients with good prognosis. In contrast, cytoplasmic-restricted CASZ1 staining or low nuclear CASZ1 staining is found in tumor samples from patients with poor prognosis. These findings provide insight into mechanisms by which CASZ1 regulates transcription, and suggests that regulation of CASZ1 subcellular localization may impact its function in normal development and pathologic conditions such as NB tumorigenesis.


Metabolic alternations in SOD-deficient Escherichia coli cells when cultivated under oxidative stress from photoexcited titanium dioxide.
Superoxide dismutase (SOD)-deficient Escherichia coli was cultivated under the oxidative stress generated by photoexcited titanium dioxide. These cells showed higher growth rate and glucose consumption rate with accelerated accumulation of acetic acid in the medium, compared to the cells cultivated under the normal condition without the stress. Under the stress condition, the activity of acetate kinase and mRNA expressions of the enzymes for acetic acid production (pta and ackA) were approximately doubled, while the activity of citrate synthase and mRNA expressions of the enzymes in TCA cycle (gltA, acnA, icd, sucA, sucC, sdhA, fumA and mdh) were repressed by about half, as compared with those under the normal condition. These results suggest that the stress-suffering cells switch the metabolic pathway into a "suppressed aerobiosis", possibly for lowering the generation of reactive oxygen species.


Interstitial Boron Atoms in the Palladium Lattice of an Industrial Type of Nanocatalyst: Properties and Structural Modifications.
It is well-established that the inclusion of small atomic species such as boron (B) in powder metal catalysts can subtly modify catalytic properties, and the associated changes in the metal lattice imply that the B atoms are located in the interstitial sites. However, there is no compelling evidence for the occurrence of interstitial B atoms, and there is a concomitant lack of detailed structural information describing the nature of this occupancy and its effects on the metal host. In this work, we use an innovative combination of high-resolution 11B magic-angle-spinning (MAS) and 105Pd static solid-state NMR nuclear magnetic resonance (NMR), synchrotron X-ray diffraction (SXRD), in situ X-ray pair distribution function (XPDF), scanning transmission electron microscopy-annular dark field imaging (STEM-ADF), electron ptychography, and electron energy loss spectroscopy (EELS) to investigate the B atom positions, properties, and structural modifications to the palladium lattice of an industrial type interstitial boron doped palladium nanoparticle catalyst system (Pd-intB/C NPs). In this study, we report that upon B incorporation into the Pd lattice, the overall face centered cubic (FCC) lattice is maintained; however, short-range disorder is introduced. The 105Pd static solid-state NMR illustrates how different types (and levels) of structural strain and disorder are introduced in the nanoparticle history. These structural distortions can lead to the appearance of small amounts of local hexagonal close packed (HCP) structured material in localized regions. The short-range lattice tailoring of the Pd framework to accommodate interstitial B dopants in the octahedral sites of the distorted FCC structure can be imaged by electron ptychography. This study describes new toolsets that enable the characterization of industrial metal nanocatalysts across length scales from macro- to microanalysis, which gives important guidance to the structure-activity relationship of the system.


Longitudinal Trends of Vascular Flow and Growth in Patients Undergoing Fontan Operation.
Single ventricle (SV) patients undergo multiple surgeries with subsequent changes in anatomy and hemodynamics. There are little cardiac magnetic resonance (CMR) data on serial changes in these patients. This study aimed to assess longitudinal changes of SV anatomy and hemodynamics in a large cohort. Anatomy and flow in SV patients with serial CMRs performed between 2008 and 2019 at a single institution were retrospectively reviewed. Mixed-effects linear regression was used to estimate changes over time at 3 to 9 months, 1 to 5 years, and >5 years after Fontan. A total of 119 patients were included (51% with hypoplastic left heart syndrome; 77% underwent extracardiac Fontan). A total of 88 patients had 3 serial CMRs. Indexed right superior vena cava, inferior vena cava, neoaortic valve, and descending aorta area decreased over time (beta = -0.19, -0.44, and -0.23, respectively; P < .01), as did indexed right superior vena cava, neoaorta and native aorta, and descending aorta flow (beta = -0.49, -0.53, and -0.59, respectively; P < .0001). Inferior vena cava flow and its contribution to total caval flow increased (beta = 0.33; P < .0001). Indexed right and left pulmonary artery flow did not change; however, indexed left pulmonary artery area decreased (beta = -0.16; P = .0014) with time. Systemic-to-pulmonary collateral flow remained unchanged before and early after Fontan (beta = -0.54; P = .42) but decreased with time from Fontan (beta = -0.22; P < .0001). In this cohort of longitudinally followed SV patients, there are significant trends in vascular size and flow over time from Fontan. These findings can be used as a framework to interpret serial CMR data in the SV and noninvasively identify deviations from expected patterns before the development of clinical symptoms.


360-Degree laser retinopexy in primary vitrectomy for rhegmatogenous retinal detachment: factors associated with its use and impact on surgical outcomes.
To determine patient and surgical factors associated with the use of 360-degree laser retinopexy during primary pars plana vitrectomy (PPV) ± scleral buckle (SB) for rhegmatogenous retinal detachment (RRD) and its impact on surgical outcomes. Patients who underwent PPV ± SB for repair of non-complex RRD at a single centre were included in this retrospective study. The primary outcome was single surgery anatomical success (SSAS). Secondary outcomes included visual acuity, epiretinal membrane formation, the presence of cystoid macular oedema, tonic pupil and corneal epithelial defects. Multiple logistic regression and multivariate regression was used. The study included 192 cases, of which 130 received 360-degree laser. Worse preoperative logMAR visual acuity (P = 0.009), male sex (P = 0.060), higher PVR grades, supplemental SB (P = 0.0468) and silicone oil/C3F8 tamponade (P < 0.0001) were associated with 360-degree laser use. No significant associations between 360-degree laser and SSAS (P = 0.079), final logMAR visual acuity (P = 0.0623), ERM development (P = 0.8208), postoperative CMO (P = 0.5946), tonic pupil (P > 0.9999) or corneal epithelial defects (P = N/A) were found. 360-degree laser retinopexy during primary PPV ± SB for RRD was associated with more complex cases and more extensive operations. Even when accounting for this, there was no difference in surgical outcomes or complication rates.


Effects of pH on the stability of the indium-113m blood protein complex and the selective binding of indium-113m to transferrin.
Indium-113m (t1/2 = 100 min; gamma-emission of 393 keV) in trace amounts was injected i.v. in rats. Blood was collected by heart puncture 15 min after the injection, and blood plasma was separated by centrifugation. Gel filtration of plasma on Sephadex G-25M equilibrated with glycine/HCl (pH 2.2-3.6), NaHCO3/CO2 (pH 4.0-11.0) glycine/NaOH (pH 8.6-10.6) or sodium acetate/acetic acid (pH 3.0-5.0) was used to separate free indium from indium bound to macromolecular proteins. Determination of radioactivity in eluted fractions showed that more than 85% of the plasma indium was bound to macromolecules at pH values between 5.0 and pH 10.6. However, dissociation of the indium plasma protein complexes occurred at pH values below 5.5, and more than 90% of the indium radioactivity was found in the low molecular weight fraction at pH 2.2. Affinity chromatography using immobilized antibodies to rat transferrin was used to isolate transferrin at pH 7.4 and 5.5. Immunodiffusion and electrophoresis were used to identify the proteins in fractions obtained by affinity chromatography. It was found that the indium-113m activity was correlated with the content of transferrin and that 80%-90% of this activity was found in fractions that had affinity to antitransferrin. These fractions contained transferrin exclusively at pH 7.4, but additional protein fractions of albumin and alpha1-globulin mobility at pH 5.5. At pH 7.4 and 5.5, 10%-20% of the indium activity was detected in molecular fractions that had no affinity to antitransferrin. Immunologic analyses showed that these fractions contained transferrin.(ABSTRACT TRUNCATED AT 250 WORDS)


Tenofovir-disoproxil-fumarate modulates lipid metabolism via hepatic CD36/PPAR-alpha activation in hepatitis B virus infection.
Entecavir and tenofovir-disoproxil-fumarate are first-line nucleos(t)ide analogs (NA) for treatment of hepatitis B virus (HBV) infections; however, their long-term administration can impact extrahepatic organs. Herein, we sought to examine the effect of NA on lipid metabolism while also characterizing the associated mechanism. A retrospective study was performed on HBV patients administered entecavir or tenofovir-disoproxil-fumarate. Patient clinical information, as well as their preserved serum samples obtained at baseline and 6-12 months after treatment initiation, were analyzed. A 1:1 propensity score matching was applied to the assignment of tenofovir-disoproxil-fumarate or entecavir treatment. Changes in serum cholesterol, including oxidized-LDL, were analyzed. Subsequently, in vitro analysis elucidated the mechanism associated with the effect of NAs on lipid metabolism. Administration of tenofovir-disoproxil-fumarate, not entecavir, to chronic HBV patients, decreased serum cholesterol levels, including non-HDL and oxidized-LDL, which are strongly associated with arteriosclerosis. In vitro analysis revealed that tenofovir-disoproxil-fumarate reduced supernatant cholesterol, and upregulated the scavenger receptor, CD36, in hepatocytes. Meanwhile, silencing of hepatic CD36 increased supernatant cholesterol and negated the cholesterol-reducing effect of tenofovir-disoproxil-fumarate in HepG2-cells. Reporter, microarray, and RT-PCR analyses further revealed that tenofovir-disoproxil-fumarate treatment activates PPAR-α-mediated signaling, and upregulates PPAR-α target genes, including CPT1 and CD36. Alternatively, silencing of PPAR-α reversed the effects of tenofovir-disoproxil-fumarate on CD36. Tenofovir-disoproxil-fumarate modulates lipid metabolism by upregulating hepatic CD36 via PPAR-α activation. Since dyslipidemia could be associated with arteriosclerosis and hepatocarcinogenesis, these discoveries provide novel insights into anti-HBV therapies, as well as the associated extrahepatic effects of NA.


Functional mapping of conserved, surface-exposed charges of antibody variable domains.
Surface-exposed charges can affect protein structure, stability and solubility as well as the kinetics of both the folding process and interaction with binding partners. We have investigated the influence on kinetic interaction parameters of 14 conserved, surface-exposed charges located away from the paratope in the variable domains of two antibodies of different specificity. We found that conserved, surface-exposed, charged framework residues are asymmetrically distributed on opposite faces of both VH and VL domains. Some of the charges play a critical role in protein folding and stability. While electrostatic forces within or close to the binding interface can be used to optimize the association rate, we confirmed the predicted minor effects of charge modifications remote from the binding site. They had no effect on the dissociation rate parameter. Our study demonstrates the role of residues remote from the interaction site in the recognition function as well as the limited effect of surface charge modifications in antibody fragments on kinetic interaction parameters.


PINK1 controls mitochondrial localization of Parkin through direct phosphorylation.
PTEN-induced putative kinase 1 (PINK1) and Parkin, encoded by their respective genes associated with Parkinson's disease (PD), are linked in a common pathway involved in the protection of mitochondrial integrity and function. However, the mechanism of their interaction at the biochemical level has not been investigated yet. Using both mammalian and Drosophila systems, we here demonstrate that the PINK1 kinase activity is required for its function in mitochondria. PINK1 regulates the localization of Parkin to the mitochondria in its kinase activity-dependent manner. In detail, Parkin phosphorylation by PINK1 on its linker region promotes its mitochondrial translocation, and the RING1 domain of Parkin is critical for this occurrence. These results demonstrate the biochemical relationship between PINK1, Parkin, and the mitochondria and thereby suggest the possible mechanism of PINK-Parkin-associated PD pathogenesis.


Toxicological study of the primaquine phosphate loaded chitosan nanoparticles in mice.
Primaquine (PQ) shows activity against the late hepatic stages and latent tissue forms of Plasmodium vivax and Plasmodium ovale. However, liver targeted PQ delivery may be useful to minimize the dose-limiting blood toxicities and side-effects of PQ. The prime objective of this study was the preparation of PQ loaded chitosan nanoparticles (PQ-CS-NPs) in order to enhance drug tolerance and to reduce dose frequency. The morphological analysis of the chitosan NPs displayed particle size in the range 287-686 nm, polydispersity index in the range 0.338-0.430 and zeta potential between 9.21 and 22.80 mV which indicated good stability. PQ-CS-NPs exhibited EE and LC as 64.28 ± 1.85% and 33.18 ± 0.975%, respectively. The in vitro drug release (Batch C7) was 97.80 ± 0.65% after 24 h. After intravenous injection of PQ-CS-NPs in mice, the lethal dose of the PQ significantly reduced when compared to that of free PQ solution.


Late physical and functional effects of osteoporotic fracture in women: the Rancho Bernardo Study.
To examine the associations between osteoporotic fractures and difficulty performing selected physical and functional activities. Cross sectional analysis of a cohort study. Geographically defined cohort located in Rancho Bernardo, California. Community-dwelling women aged 55 and older who participated in a study of osteoporosis between 1988-1991. Eighty percent of eligible women participated in the study. Self-reported difficulty performing seven physical activities and four functional tasks. The mean age of the 1010 women was 72.6 years. A total of 160 first minimal trauma fractures occurred between 1972 and 1991, including 62 wrist, 29 rib, 25 hip, and 23 spine fractures. The mean time since fracture was 6.7 years (range, 1 to 17 years). In multiply adjusted analyses, having experienced any osteoporotic fracture was significantly associated with a 1.7 to 3.0-fold increase in difficulty bending, lifting, reaching, walking, climbing stairs, and descending stairs. Any fracture was significantly associated with 1.9 to 6.7 times more difficulty in dressing, cooking, shopping, and performing heavy housework. Compared with the relative odds of physical limitation associated with any osteoporotic fracture, hip fractures were more strongly associated with difficulty walking (OR 3.6) and descending stairs (OR 4.1), whereas spine fractures demonstrated a stronger association with difficulty bending (OR 3.1), lifting (OR 3.4), and descending stairs (OR 4.2). Among older women, remote osteoporotic fracture at any site is associated with an approximate doubling of the risk of physical limitations and an even higher risk of functional limitations. Although this cross-sectional analysis cannot secure the direction of the association, the specificity of the effect of particular fractures on discrete activities supports causality.


Comparison of the complement-fixation and agar gel immunodiffusion tests for diagnosis of subclinical bovine paratuberculosis.
The performance of the serum complement fixation (CF) test was compared with that of a serum agar gel immunodiffusion (AGID) test on 74 subclinically infected and 154 uninfected cattle in 6 commercial midwestern dairy herds with Mycobacterium paratuberculosis infection and on 30 cattle in a herd that was free of infection. Infection status of cattle within herds was established by performance of a series of 3 or more fecal cultures and of ileocecal lymph node cultures of culled cattle. In cattle with subclinical infection detected by culturing, the sensitivity estimates of the CF and AGID tests were 10.8% (3.6% SE) and 18.9% (4.5% SE), respectively. In the cattle classified as disease free, the specificity estimates of the CF and AGID tests were 97.4% (1.3% SE) and 99.4% (0.6% SE), respectively. Neither set of estimates was significantly different. Negative test results obtained with the use of either test in apparently normal cattle from suspect herds should be interpreted with caution because both tests suffer from low sensitivities in subclinically infected animals. However, the AGID test may be more useful in regulatory situations in which the CF test is currently used because the AGID test is easier to perform and to interpret.


The significance of the three volar spaces in forearm compartment syndrome: a clinical and cadaveric correlation.
The goal of this study was to assess the importance of the 3 volar spaces in forearm compartment syndrome in a prospective manner. A cadaveric model was developed to correlate with our clinical experience. All but 1 of 21 volar compartments (superficial volar, deep volar, and pronator quadratus spaces) in 7 patients in our clinical series decompressed adequately after release of the superficial volar fascia. One patient needed further release of the pronator quadratus compartment; he had suffered a combined crush and vascular injury. All volar compartments in our cadaveric model decompressed with superficial fasciotomy. Prefasciotomy and postfasciotomy pressures should be obtained from all 3 compartments of the volar forearm. Superficial fasciotomy usually adequately decompresses the entire volar forearm; however, in the event that deep compartment pressures remain high after superficial fasciotomy, release of the affected space is indicated.


Analysis of the factor XI variant Arg184Gly suggests a structural basis for factor IX binding to factor XIa.
A patient with factor XI (FXI) deficiency was reported with an Arg184Gly substitution in the FXI A3 domain. The A3 domain contains an exosite required for binding of FIX to activated FXI (FXIa). To test the effects of the Arg184Gly substitution on FIX activation, and to characterize the FIX-binding site on FXIa. Recombinant FXIa and FIX variants were used to identify residues involved in FIX activation by FXIa. Analysis of the FXI structure was used to identify potential FIX-binding sites. The Km for FIX activation by FXIa-Gly184 was approximately three-fold higher than for FXIa, suggesting that Arg184 is part of the exosite. Arg184 and the adjacent residues, Ile183 and Asp185, contribute to charged and hydrophobic areas that are not present in the FXI homolog prekallikrein (PK). Replacing residues 183-185 with alanine abolished exosite activity, similarly to replacement of the entire A3 domain with the A3 domain from PK (FXIa/PKA3). Reintroducing FXI residues 183-185 into FXIa/PKA3 partially restored the exosite, and replacing residues 183-185 and 260-264 completely restored exosite function. FIX in which the Ω-loop (residues 4-11) was replaced with the FVII Ω-loop was activated poorly by FXIa, suggesting that the FIX Ω-loop binds to FXIa. The results support a model in which the Ω-loop of FIX binds to an area on FXIa composed of residues from the N-terminus and C-terminus of the A3 domain. These residues are buried in zymogen FXI, and must be exposed upon conversion to FXIa to permit FIX binding.