Interaction studies of multiple binding sites on m4 muscarinic acetylcholine receptors.
This study investigated the reciprocal cross-interactions between two distinct allosteric sites on the M(4) muscarinic acetylcholine receptor (mAChR) in the absence or presence of different orthosteric ligands. Initial studies revealed that two novel benzimidazole allosteric modulators, 17-beta-hydroxy-17-alpha-ethy nyl-delta(4)-androstano[3,2-b]pyrimido[1,2-a]benzimidazole (WIN 62,577) and 17-beta-hydroxy-17-alpha-ethynyl-5-alpha-androstano[3,2-b]pyrimido[1,2-a]benzimidazole (WIN 51,708), exhibited different degrees of positive, negative, or close-to-neutral cooperativity with the orthosteric site on M(1) or M(4) mAChRs, depending on the chemical nature of the orthosteric radioligand that was used [[(3)H]N-methylscopolamine ([(3)H]NMS) versus [(3)H]quinuclidinylbenzilate ([(3)H]QNB)]. The largest window for observing an effect (negative cooperativity) was noted for the combination of WIN 62,577 and [(3)H]QNB at the M(4) mAChR. Experiments involving the combination of these two ligands with unlabeled agonists [acetylcholine, 4-(m-chlorophenylcarbamoyloxy)-2-butynyltrimethylammonium (McN-A-343), or xanomeline] revealed low degrees of negative cooperativity between WIN 62,577 and each agonist, whereas stronger negative cooperativity was observed against atropine. It is interesting that when these experiments were repeated using the prototypical modulators heptane-1,7-bis-(dimethyl-3'-phthalimidopropyl)-ammonium bromide (C(7)/3-phth), alcuronium, or brucine (which act at a separate allosteric site), WIN 62,577 exhibited negative cooperativity with each modulator when the orthosteric site was unoccupied, but this switched to neutral cooperativity when the receptor was occupied by [(3)H]QNB. Dissociation kinetic experiments using [(3)H]NMS and combination of C(7)/3-phth with WIN 62,577 also provided evidence for neutral cooperativity between the two allosteric sites when the orthosteric site is occupied. Together, these results provide insight into the nature of the interaction between two distinct allosteric sites on the M(4) mAChR and how this interaction is perturbed upon occupancy of the orthosteric site.


NAD+-induced vasotoxicity in the pericyte-containing microvasculature of the rat retina: effect of diabetes.
It was recently proposed that activation of P2X(7) purinoceptors may play a role in causing cell death in the pericyte-containing microvasculature of the diabetic retina. This hypothesis is supported by the observation that diabetes enhances lethal pore formation in retinal microvessels exposed to synthetic P2X(7) agonists. The goal of this study was to determine whether purinergic vasotoxicity can be triggered by the endogenous molecule nicotinamide adenosine dinucleotide (NAD(+)), which is a substrate for ecto-ribosylation reactions known to activate P2X(7) receptor/channels in other cell types. Pericyte-containing retinal microvessels were isolated from normal and streptozotocin-injected rats. Trypan blue dye exclusion was used to assess cell viability, YO-PRO-1 uptake was used to identify cells with P2X(7)-induced pores, and ethenoadenosine antibodies were used to detect ecto-adenosine diphosphate (ADP)-ribosyltransferase (ART) activity. In freshly isolated retinal microvessels, it was found that extracellular NAD(+), but not its catabolites, caused cell death (half-maximal effective concentration [EC(50)] = 2 nM) by a mechanism involving the activation of P2X(7) purinoceptors and the formation of transmembrane pores. A series of experiments provided evidence that NAD(+), which is not a direct purinergic agonist, serves as a substrate for ecto-ribosylation reactions that subsequently trigger P2X(7)-dependent cell death in the retinal microvasculature. Soon after the onset of diabetes, the sensitivity of retinal microvessels to the vasotoxic effect of extracellular NAD(+) increased by approximately 100-fold. Purinergic vasotoxicity triggered by extracellular NAD(+) is a newly recognized mechanism that may contribute to the cell death observed in the pericyte-containing microvascular of the diabetic retina.


High frequency of cross-reactive cytotoxic T lymphocytes elicited during the virus-induced polyclonal cytotoxic T lymphocyte response.
Polyclonal stimulation of CD8+ cytotoxic T lymphocytes (CTL) occurs during infection with many viruses including those not known to transform CTL or encode superantigens. This polyclonal CTL response includes the generation of high levels of allospecific CTL directed against many class I haplotypes. In this report we investigated whether the allospecific CTL generated during an acute lymphocytic choriomeningitis virus (LCMV) infection of C57BL/6 mice were stimulated specifically by antigen recognition or nonspecifically by polyclonal mechanisms possibly involving lymphokines or superantigens. An examination of the ability of different strains of mice to induce high levels of CTL specific for a given alloantigen showed that most, but not all, strains generated high levels of allospecific CTL, and that their abilities to generate them mapped genetically to the major histocompatibility complex locus, exclusive of the class II region. This indicated that the virus-induced allospecific CTL generation was independent of the class II allotype, and mice depleted of CD4+ cells generated allospecific CTL, indicating independence of class II-CD4+ cell interactions and resulting CD4+ cell-secreted lymphokines. FACS staining with a variety of V beta-binding antibodies did not show a superantigen-like depletion or enrichment of any tested V beta + subset during infection. Several experiments provided evidence in support of direct stimulation of CD8+ cells via the T cell receptor: (a) both virus- and allo-specific killing were enriched within a given V beta subpopulation; (b) relative CTL precursor frequencies against different class I alloantigens changed during the course of virus infection; (c) the relative levels of virus-induced, allospecific CTL-mediated lysis at day 8 after infection did not parallel the CTL precursor frequencies before infection; and (d) limiting dilution analyses of day 8 LCMV-infected spleen cells stimulated by virus-infected syngeneic peritoneal exudate cells (PEC) revealed not only the expected virus-specific CTL clones, but also a high frequency of clones that were cross-reactive with allogeneic and virus-infected syngeneic targets. In addition to the virus cross-reactive allospecific CTL clones, virus-infected PEC also stimulated the generation of some allospecific clones that did not lyse virus-infected fibroblasts. Surprisingly, LCMV-infected PEC were much more efficient at stimulating allospecific CTL clones from day 8 LCMV-infected splenocytes than were allogeneic stimulators. These results indicate that at least part of the polyclonal allospecific CTL response elicited by acute virus infection is a consequence of the selective expansion of many clones of allospecific CTL which cross-react with virus-infected cells.(ABSTRACT TRUNCATED AT 400 WORDS)


[Plasmacytoid dendritic cells: the novel Eldorado for antiviral therapy?].
Plasmacytoid dendritic cells (pDCs) represent the first line of host defense against viruses and are an essential link between innate and adaptive immunity. The antiviral factor IFN-α is massively produced by pDCs in response to HIV infection and induces the expression of cellular genes that interfere with viral replication (ISG). Indeed, type I IFN produced by pDCs has a direct anti-viral activity against HIV and has important adjuvant function on other immune cell-types, such as T cells, macrophages and dendritic cells. However, the role of type I IFN in HIV disease is complex and may depend on the stage of the disease. The immunologic hallmark of HIV infection is a status of chronic and progressive immune activation, which drives the immune system to exhaustion and leads to severe immunodeficiency. There is now strong evidence that chronic activation of pDCs may promote HIV pathogenesis and have an impact on adaptive T-cell response. Thus, targeting pDCs and type I IFN may open new therapeutic strategies for chronically activated HIV patients.


Cysteine-modified orange peel for removal of Cu(II) from aqueous solutions.
In the present work, cysteine-modified orange peel (COP) for the removal of Cu(II) from aqueous solutions has been developed and comparatively studied with diethylenetriamine-modified orange peel (DOP). Both COP and DOP were systematically evaluated via their capabilities for adsorbing Cu(II), including the key influential parameters such as initial pH, contact time and initial Cu(II) concentration. Further studies suggest that the sorption of Cu(II) onto both COP and DOP fits well with the pseudo-second-order equation, and the corresponding sorption isotherm can be classified to a Langmuir isotherm model. COP appears more advantageous over DOP and far better than that of unmodified OP in removal of Cu(II) from aqueous system. The maximum capacities of COP and DOP for adsorbing Cu(II) are 95.23 and 83.68 mg/g, respectively, about three times higher than that of unmodified OP. The sorption efficiency of COP drops by merely about 3% after five cycles, implying a promising usage in the removal of Cu(II) from wastewater in practice.


Peierls-Nabarro energy surfaces and directional mobility of discrete solitons in two-dimensional saturable nonlinear Schrödinger lattices.
We address the problem of directional mobility of discrete solitons in two-dimensional rectangular lattices, in the framework of a discrete nonlinear Schrödinger model with saturable on-site nonlinearity. A numerical constrained Newton-Raphson method is used to calculate two-dimensional Peierls-Nabarro energy surfaces, which describe a pseudopotential landscape for the slow mobility of coherent localized excitations, corresponding to continuous phase-space trajectories passing close to stationary modes. Investigating the two-parameter space of the model through independent variations of the nonlinearity constant and the power, we show how parameter regimes and directions of good mobility are connected to the existence of smooth surfaces connecting the stationary states. In particular, directions where solutions can move with minimum radiation can be predicted from flatter parts of the surfaces. For such mobile solutions, slight perturbations in the transverse direction yield additional transverse oscillations with frequencies determined by the curvature of the energy surfaces, and with amplitudes that for certain velocities may grow rapidly. We also describe how the mobility properties and surface topologies are affected by inclusion of weak lattice anisotropy.


High temporal frequency synchrony is insufficient for perceptual grouping.
We used textures of randomly moving grating patches to assess the role of fine-grain temporal synchrony in texture segregation. In the target area, patches reversed direction simultaneously. In the surround, patches changed direction at random times. Thus, phase changes in the target area were precisely synchronous, whereas those in the surround were not. In agreement with work carried out by Lee and Blake, we found that the target area was frequently visible, and that observers could discriminate its shape (horizontal versus vertical) at frame rates of 100 Hz in brief exposures (200 ms). Further experiments suggested that the length of unidirectional motion sequences in the target area, rather than synchrony, determined its visibility. To eliminate completely contrast and motion cues, we made all the background elements identical to the target elements, but with a random starting phase. Despite the presence of synchrony in the target area but not the background, the target was generally very hard to see. Targets that remained visible contained low temporal frequency modulations of direction. We conclude that the human observer can detect synchrony, but only at modest temporal frequencies once motion and contrast artefacts have been eliminated.


Synthesis of oxazolidinones: rhodium-catalyzed C-H amination of N-mesyloxycarbamates.
N-Mesyloxycarbamates undergo intramolecular C-H amination reactions to afford oxazolidinones in good to excellent yields in the presence of rhodium(ii) carboxylate catalysts. The reaction is performed under green conditions and potassium carbonate is used, forming biodegradable potassium mesylate as a reaction by-product. This method enables the production of electron-rich, electron-deficient, aromatic and heteroaromatic oxazolidinones in good to excellent yields. Conformationally restricted cyclic secondary N-mesyloxycarbamates furnish cis-oxazolidinones in high yields and selectivity; DFT calculations are provided to account for the observed selectivity. trans-Oxazolidinones were prepared from acyclic secondary N-mesyloxycarbamates using Rh2(oct)4. The selectivity was reverted with a cytoxazone N-mesyloxycarbamate precursor using large chiral rhodium(ii) carboxylate complexes, affording the corresponding cis-oxazolidinone. This orthogonal selectivity was used to achieve the formal synthesis of (-)-cytoxazone.


Light composite scalar in twelve-flavor QCD on the lattice.
On the basis of lattice simulations using highly improved staggered quarks for twelve-flavor QCD with several bare fermion masses, we observe a flavor-singlet scalar state lighter than the pion in the correlators of fermionic interpolating operators. The same state is also investigated using correlators of gluonic interpolating operators. Combined with our previous study that showed twelve-flavor QCD to be consistent with being in the conformal window, we infer that the lightness of the scalar state is due to infrared conformality. This result shed some light on the possibility of a light composite Higgs boson ("technidilaton") in walking technicolor theories.


Minimal residual disease after allogeneic bone marrow transplantation for chronic myeloid leukaemia: a metaphase-FISH study.
Metaphase-FISH was adopted for the detection of proliferating Philadelphia-positive (Ph+) residual leukaemic cells in 25 patients with chronic myeloid leukaemia treated with allogeneic bone marrow transplantation (BMT). Patients were followed up during their clinical remission for 4-50 months (median 17 months) after BMT. 80 bone marrow samples were studied. For most of the cases no fewer than 1000 metaphases were analysed. Six patients (24%) showed residual Ph+ cells during the first 6 months and two others by the end of the first year after BMT. Three patients relapsed during the study and in two of them residual Ph+ cells were detected during the first 6 months after BMT. In 17 patients no Ph+ cells were detected at any stage of follow-up and 16 (94.1%) of them continue in complete clinical and haematological remission. Our results indicate that metaphase-FISH is a reliable tool in the quantitation of proliferating residual leukaemic cells. We suggest that consecutive findings of equal amounts of residual leukaemic cells do not necessarily predict a relapse. However, their presence calls for follow-up at shorter intervals where an increasing number of these cells predicts an ensuing relapse.


GIS applications in land management: the loss of high quality land to development in Central Mississippi from 1987-2002.
The socio-economic trends and history of Central Mississippi reveal a major rural influence based upon a dependence on agricultural activities as part of the economic engine driving the state's economy. Yet, in the last several years, the amount of agricultural land in the counties continues to decline. Similar changes in other variables associated with agricultural land use and the continuity of farming in the state have also been changing. Indeed, under the pressure of urban growth, some farmers are forced to use less productive soils or have abandoned the agricultural business. Considering the gravity of the problem and the implications for sustainable development, public concern has increased in the state of Mississippi that urbanization and other factors may be eroding potential farmland. Given the effects of the current trends on the future capacity to produce food items, there are concerns that the growing incidence of farmland loss may also erode the basis for sustainable use of agricultural land, biodiversity and protection of the state's ecological treasures. Notwithstanding the gravity of these trends, no major effort in the literature has aimed at documenting the incidence of agricultural land loss and the linkages to urbanization in the region of Central Mississippi. What changes have taken place in the size of agricultural land within the counties and what factors are responsible for it? This paper examines the issue of farmland loss in Central Mississippi with a focus at the county level between 1987 and 2002 from a temporal-spatial perspective. In terms of methodology, the paper uses a mixed scale approach based upon the existing literature. Data were drawn from the United States Census databases of Population and Agriculture. This information is analyzed with basic descriptive statistics and GIS with particular attention to the spatial trends at the county level. Results indicate that the counties under consideration have experienced considerable change in the amount of agricultural land and other variables associated with the use of farmland, due to urbanization. With the types of changes occurring, instituting effective policies anchored in sustainability, community participation, and growth management will go a long way in addressing the situation. Other strategies for farmland protection based upon land information inventory and mapping in the region, are also recommended. The paper stands as an update of the existing literature and offers a valuable tool for decision makers within the domain of natural resources management.


Stigmatization and self-determination of preregistration nurses: A path analysis.
The concept of stigma and the stigmatizing behaviours of health-care professionals can have a profound influence on people with mental illness. A key construct that has been identified as influencing our behaviours is self-determination. As such, in the present study, we attempted to examine the connection and influence of motivational measures on the stigmatization of preregistration nurses. Data were collected once using three surveys that measured the motivational responses and stigmatizing behaviours of preregistration nurses after an approved mental health clinical placement. Using a path analysis, the results indicated that psychological needs significantly influenced preregistration nurses' self-determination towards work. In addition, self-determination was a significant influence on the stigmatization behaviours of preregistration nurses. The results of the present study provide initial empirical evidence that supports the importance of professional self-determination and the potential connection of quality care, as illustrated with the low stigmatizing behaviours of preregistration nurses who are more self-determined towards their work/career. Because of the significant results of the present study, it is recommended that future research is needed that uses self-determination theory as a lens to understand the application and importance within the field of nurse education.


Breast cancer cell-derived fibroblast growth factors enhance osteoclast activity and contribute to the formation of metastatic lesions.
Fibroblast growth factors (FGFs) and their receptors (FGFRs) have been implicated in promoting breast cancer growth and progression. While the autocrine effects of FGFR activation in tumor cells have been extensively studied, little is known about the effects of tumor cell-derived FGFs on cells in the microenvironment. Because FGF signaling has been implicated in the regulation of bone formation and osteoclast differentiation, we hypothesized that tumor cell-derived FGFs are capable of modulating osteoclast function and contributing to growth of metastatic lesions in the bone. Initial studies examining FGFR expression during osteoclast differentiation revealed increased expression of FGFR1 in osteoclasts during differentiation. Therefore, studies were performed to determine whether tumor cell-derived FGFs are capable of promoting osteoclast differentiation and activity. Using both non-transformed and transformed cell lines, we demonstrate that breast cancer cells express a number of FGF ligands that are known to activate FGFR1. Furthermore our results demonstrate that inhibition of FGFR activity using the clinically relevant inhibitor BGJ398 leads to reduced osteoclast differentiation and activity in vitro. Treatment of mice injected with tumor cells into the femurs with BGJ398 leads to reduced osteoclast activity and bone destruction. Together, these studies demonstrate that tumor cell-derived FGFs enhance osteoclast function and contribute to the formation of metastatic lesions in breast cancer.


Occupational physical demand and risk of hip fracture in older women.
Hip fractures are leading causes of disability, morbidity and mortality among older women. Since physical activity helps maintain physical functioning and bone mineral density, occupational physical demand may influence fracture risk. This study investigates the association of occupational physical demand with hip fracture incidence among women. The Women's Health Initiative Observational Study is a multiethnic cohort of 93,676 postmenopausal women, 50-79 years of age at enrolment, enrolled from 1994 to 1998 at 40 geographically diverse clinical centres throughout the USA. Outcomes including hip fractures were assessed annually and up to 3 jobs held since age 18 years were reported by each woman. Occupational physical demand levels were assigned for each job through linkage of occupational titles with Standard Occupational Codes and the Occupational Information Network. Average, cumulative and peak physical demand scores both before and after menopause and throughout women's work life were estimated. Women were followed through 2010 for an average of 11.5 years; 1834 hip fractures occurred during this time. We did not observe an overall association of occupational physical demand with subsequent risk of hip fracture after adjusting for age, race/ethnicity, birth region and education. Previous research on occupations and hip fracture risk in women is inconclusive. This study was able to take critical risk periods into account and control for confounding factors in a large cohort of older women to show that overall occupational physical demand neither increases nor decreases risk of hip fracture later in life.


Is secondary preventive care improving? Observational study of 10-year trends in emergency admissions for conditions amenable to ambulatory care.
To identify trends in emergency admissions for patients with clinical conditions classed as 'ambulatory care sensitive' (ACS) and assess if reductions might be due to improvements in preventive care. Observational study of routinely collected hospital admission data from March 2001 to April 2011. Admission rates were calculated at the population level using national population estimates for area of residence. All emergency admissions to National Health Service (NHS) hospitals in England from April 2001 to March 2011 for people residents in England. Age-standardised emergency admissions rates for each of 27 specific ACS conditions (ICD-10 codes recorded as primary or secondary diagnoses). Between April 2001 and March 2011 the number of admissions for ACS conditions increased by 40%. When ACS conditions were defined solely on primary diagnosis, the increase was less at 35% and similar to the increase in emergency admissions for non-ACS conditions. Age-standardised rates of emergency admission for ACS conditions had increased by 25%, and there were notable variations by age group and by individual condition. Overall, the greatest increases were for urinary tract infection, pyelonephritis, pneumonia, gastroenteritis and chronic obstructive pulmonary disease. There were significant reductions in emergency admission rates for angina, perforated ulcers and pelvic inflammatory diseases but the scale of these successes was relatively small. Increases in rates of emergency admissions suggest that efforts to improve the preventive management of certain clinical conditions have failed to reduce the demand for emergency care. Tackling the demand for hospital care needs more radical approaches than those adopted hitherto if reductions in emergency admission rates for ACS conditions overall are to be seen as a positive outcome of for NHS.


Pruriceptive spinothalamic tract neurons: physiological properties and projection targets in the primate.
Itch of peripheral origin requires information transfer from the spinal cord to the brain for perception. Here, primate spinothalamic tract (STT) neurons from lumbar spinal cord were functionally characterized by in vivo electrophysiology to determine the role of these cells in the transmission of pruriceptive information. One hundred eleven STT neurons were identified by antidromic stimulation and then recorded while histamine and cowhage (a nonhistaminergic pruritogen) were sequentially applied to the cutaneous receptive field of each cell. Twenty percent of STT neurons responded to histamine, 13% responded to cowhage, and 2% responded to both. All pruriceptive STT neurons were mechanically sensitive and additionally responded to heat, intradermal capsaicin, or both. STT neurons located in the superficial dorsal horn responded with greater discharge and longer duration to pruritogens than STT neurons located in the deep dorsal horn. Pruriceptive STT neurons discharged in a bursting pattern in response to the activating pruritogen and to capsaicin. Microantidromic mapping was used to determine the zone of termination for pruriceptive STT axons within the thalamus. Axons from histamine-responsive and cowhage-responsive STT neurons terminated in several thalamic nuclei including the ventral posterior lateral, ventral posterior inferior, and posterior nuclei. Axons from cowhage-responsive neurons were additionally found to terminate in the suprageniculate and medial geniculate nuclei. Histamine-responsive STT neurons were sensitized to gentle stroking of the receptive field after the response to histamine, suggesting a spinal mechanism for alloknesis. The results show that pruriceptive information is encoded by polymodal STT neurons in histaminergic or nonhistaminergic pathways and transmitted to the ventrobasal complex and posterior thalamus in primates.


Rheumatic fever: autoantibodies against a variety of cardiac, nuclear, and streptococcal antigens.
To measure antibody titres to cardiac, nuclear, and streptococcal antigens in different groups of rheumatic fever (n = 60) and control subjects (n = 80) with the aim of identifying cross reactive antigens of potential laboratory diagnostic value. Enzyme linked immunosorbent assays (ELISA), immunocytochemical, and electrophoretic techniques were used to measure titres of antibodies to a variety of cardiac, nuclear, and streptococcal antigens in seven groups comprising patients with rheumatic fever and control subjects. Increased concentrations of antibodies to several streptococcal and cardiac antigens, in addition to increased IgA and IgG levels, were noted in sera from patients with acute rheumatic fever and chronic rheumatic heart disease. Autoantibodies to nuclear antigens were evident in three rheumatic fever sera. Although we were unable to identify any unique cross reactivity between cardiac and streptococcal antigens, these results demonstrate that there is an exaggerated humoral response to several cardiac, nuclear and streptococcal antigens in patients with rheumatic fever.


Characteristics and outcomes of acute kidney injury in hospitalized COVID-19 patients: A multicenter study by the Turkish society of nephrology.
Acute kidney injury (AKI) is common in coronavirus disease-2019 (COVID-19) and the severity of AKI is linked to adverse outcomes. In this study, we investigated the factors associated with in-hospital outcomes among hospitalized patients with COVID-19 and AKI. In this multicenter retrospective observational study, we evaluated the characteristics and in-hospital renal and patient outcomes of 578 patients with confirmed COVID-19 and AKI. Data were collected from 34 hospitals in Turkey from March 11 to June 30, 2020. AKI definition and staging were based on the Kidney Disease Improving Global Outcomes criteria. Patients with end-stage kidney disease or with a kidney transplant were excluded. Renal outcomes were identified only in discharged patients. The median age of the patients was 69 years, and 60.9% were males. The most frequent comorbid conditions were hypertension (70.5%), diabetes mellitus (43.8%), and chronic kidney disease (CKD) (37.6%). The proportions of AKI stages 1, 2, and 3 were 54.0%, 24.7%, and 21.3%, respectively. 291 patients (50.3%) were admitted to the intensive care unit. Renal improvement was complete in 81.7% and partial in 17.2% of the patients who were discharged. Renal outcomes were worse in patients with AKI stage 3 or baseline CKD. The overall in-hospital mortality in patients with AKI was 38.9%. In-hospital mortality rate was not different in patients with preexisting non-dialysis CKD compared to patients without CKD (34.4 versus 34.0%, p = 0.924). By multivariate Cox regression analysis, age (hazard ratio [HR] [95% confidence interval (95%CI)]: 1.01 [1.0-1.03], p = 0.035], male gender (HR [95%CI]: 1.47 [1.04-2.09], p = 0.029), diabetes mellitus (HR [95%CI]: 1.51 [1.06-2.17], p = 0.022) and cerebrovascular disease (HR [95%CI]: 1.82 [1.08-3.07], p = 0.023), serum lactate dehydrogenase (greater than two-fold increase) (HR [95%CI]: 1.55 [1.05-2.30], p = 0.027) and AKI stage 2 (HR [95%CI]: 1.98 [1.25-3.14], p = 0.003) and stage 3 (HR [95%CI]: 2.25 [1.44-3.51], p = 0.0001) were independent predictors of in-hospital mortality. Advanced-stage AKI is associated with extremely high mortality among hospitalized COVID-19 patients. Age, male gender, comorbidities, which are risk factors for mortality in patients with COVID-19 in the general population, are also related to in-hospital mortality in patients with AKI. However, preexisting non-dialysis CKD did not increase in-hospital mortality rate among AKI patients. Renal problems continue in a significant portion of the patients who were discharged.


Subclinical hepatic dysfunction in laparoscopic cholecystectomy and laparoscopic colectomy.
Laparoscopic surgery causes a reduction in hepatic blood flow due to a number of factors, including raised intra-abdominal pressure, the neurohumoral response to surgical stress and the effect of patient position. The clinical significance of the phenomenon is not fully understood. Plasma concentrations of alcohol dehydrogenase (AD) and glutathione S-transferase (GST), which are concentrated in the centrilobular acinus of the liver, sensitively reflect hepatic hypoperfusion, and can be used to monitor reductions in hepatic blood flow. We compared perioperative AD, GST, aspartate aminotransferase (AST, normal range 14-32 IU litre(-1)) and alanine aminotransferase (ALT, normal range 8-41 U litre(-1)) concentrations in patients undergoing laparoscopic cholecystectomy or laparoscopic colectomy to study how patient position and surgical manipulation of the liver affect hepatocellular integrity during laparoscopy. There were significant postoperative increases in AD and GST in the cholecystectomy group [mean (SD) peak concentration 10.8 (4.7) U litre(-1) and 113 (55) microg litre(-1) respectively]. Although the duration of pneumoperitoneum was longer in the colectomy group, there were no comparable perioperative increases in AD and GST in this group [peak concentration 4.0 (4.0) U litre(-1) and 33 (35) microg litre(-1) respectively]. AST and ALT on the first postoperative day were significantly higher in the laparoscopic cholecystectomy group (41 and 34 U litre(-1) respectively) than in the laparoscopic colectomy group (24 and 18 U litre(-1); P<0.05 for each). These results indicate that patient position and the effects of surgical manipulation of the liver affect perioperative hepatic perfusion significantly.


Engineering Streptomyces coelicolor for heterologous expression of secondary metabolite gene clusters.
We have constructed derivatives of Streptomyces coelicolor M145 as hosts for the heterologous expression of secondary metabolite gene clusters. To remove potentially competitive sinks of carbon and nitrogen, and to provide a host devoid of antibiotic activity, we deleted four endogenous secondary metabolite gene clusters from S. coelicolor M145--those for actinorhodin, prodiginine, CPK and CDA biosynthesis. We then introduced point mutations into rpoB and rpsL to pleiotropically increase the level of secondary metabolite production. Introduction of the native actinorhodin gene cluster and of gene clusters for the heterologous production of chloramphenicol and congocidine revealed dramatic increases in antibiotic production compared with the parental strain. In addition to lacking antibacterial activity, the engineered strains possess relatively simple extracellular metabolite profiles. When combined with liquid chromatography and mass spectrometry, we believe that these genetically engineered strains will markedly facilitate the discovery of new compounds by heterologous expression of cloned gene clusters, particularly the numerous cryptic secondary metabolic gene clusters that are prevalent within actinomycete genome sequences.


Effect of anterior cruciate healing on the uninjured ligament insertion site.
The effect of anterior cruciate healing on the uninjured ligament insertion site after enhanced suture repair with collagen-platelet composites (CPC) has not yet been defined. In this study, we hypothesized that fibroblasts and osteoclasts would participate in generating histologic changes in insertion site morphology after transection and bioenhanced repair of the ACL, and that these changes would be age-dependent. Skeletally immature, adolescent, and adult Yucatan mini-pigs underwent ACL transection and bioenhanced suture repair. The histologic response to repair of the insertion site was evaluated at 1, 2, 4, and 15 weeks. In young and adolescent animals treated with bioenhanced suture repair with CPC, changes in the insertion site included: (1) fibroblastic proliferation with loss and return of collagen alignment in the fibrous zone; (2) osteoclastic resorption within fibrocartilage zones at 2-4 weeks; and (3) partial reappearance of fibrocartilage zones at 15 weeks. In adult animals; however, degenerative changes were noted by 15 weeks: (1) loss of parallel arrangement of collagen fibers in the fibrous zone; and (2) increasing disorganization and loss of columnation of chondrocytes in the fibrocartilage zone. These results suggest that fibroblasts and osteoclasts mediate histologic changes at the insertion site during bioenhanced suture repair of the ACL which may prevent insertion site degeneration, and that the magnitude of these changes may be a function of skeletal maturity.


Diabetic nephropathy induces alterations in the glomerular and tubule lipid profiles.
Diabetic nephropathy (DN) is a major life-threatening complication of diabetes. Renal lesions affect glomeruli and tubules, but the pathogenesis is not completely understood. Phospholipids and glycolipids are molecules that carry out multiple cell functions in health and disease, and their role in DN pathogenesis is unknown. We employed high spatial resolution MALDI imaging MS to determine lipid changes in kidneys of eNOS(-/-) db/db mice, a robust model of DN. Phospholipid and glycolipid structures, localization patterns, and relative tissue levels were determined in individual renal glomeruli and tubules without disturbing tissue morphology. A significant increase in the levels of specific glomerular and tubular lipid species from four different classes, i.e., gangliosides, sulfoglycosphingolipids, lysophospholipids, and phosphatidylethanolamines, was detected in diabetic kidneys compared with nondiabetic controls. Inhibition of nonenzymatic oxidative and glycoxidative pathways attenuated the increase in lipid levels and ameliorated renal pathology, even though blood glucose levels remained unchanged. Our data demonstrate that the levels of specific phospho- and glycolipids in glomeruli and/or tubules are associated with diabetic renal pathology. We suggest that hyperglycemia-induced DN pathogenic mechanisms require intermediate oxidative steps that involve specific phospholipid and glycolipid species.


Tyrosine phosphorylation of the insulin receptor during insulin-stimulated internalization in rat hepatoma cells.
We have studied the phosphorylation state of the insulin receptor during receptor-mediated endocytosis in the well-differentiated rat hepatoma cell line Fao. Insulin induced the rapid internalization of surface-iodinated insulin receptors into a trypsin-resistant compartment, with a 3-fold increase in the internalization rate over that seen in the absence of insulin. Within 20 min of insulin stimulation, 30-35% of surface receptors were located inside the cell. This redistribution was half-maximal by 10.5 min. Similar results were obtained when the loss of surface receptors was measured by 125I-insulin binding. Tyrosyl phosphorylation of internalized insulin receptors was measured by immunoprecipitation with antiphosphotyrosine antibody. Immediately after insulin stimulation, 70-80% of internalized receptors were tyrosine phosphorylated. Internalized receptors persisted in a phosphorylated state after the dissociation of insulin but were dephosphorylated prior to their return to the plasma membrane. After 45-60 min of insulin stimulation, the tyrosine phosphorylation of the internal receptor pool decreased by 45%, whereas the phosphorylation of surface receptors was unchanged. These data suggest that insulin induces the internalization of phosphorylated insulin receptors into the cell and that the phosphorylation state of the internal receptor pool may be regulated by insulin.


Computational modelling of efflux pumps and their inhibitors.
Antimicrobial resistance is based on the multifarious strategies that bacteria adopt to face antibiotic therapies, making it a key public health concern of our era. Among these strategies, efflux pumps (EPs) contribute significantly to increase the levels and profiles of resistance by expelling a broad range of unrelated compounds - buying time for the organisms to develop specific resistance. In Gram-negative bacteria, many of these chromosomally encoded transporters form multicomponent 'pumps' that span both inner and outer membranes and are driven energetically by a primary or secondary transporter component.One of the strategies to reinvigorate the efficacy of antimicrobials is by joint administration with EP inhibitors (EPI), which either block the substrate binding and/or hinder any of the transport-dependent steps of the pump. In this review, we provide an overview of multidrug-resistance EPs, their inhibition strategies and the relevant findings from the various computational simulation studies reported to date with respect to deciphering the mechanism of action of inhibitors with the purpose of improving their rational design.


Gastrointestinal injury associated with NSAID use: a case study and review of risk factors and preventative strategies.
Nonsteroidal anti-inflammatory drugs (NSAIDs) are effective anti-inflammatory and analgesic agents and are among the most commonly used classes of medications worldwide. However, their use has been associated with potentially serious dose-dependent gastrointestinal (GI) complications such as upper GI bleeding. GI complications resulting from NSAID use are among the most common drug side effects in the United States, due to the widespread use of NSAIDs. The risk of upper GI complications can occur even with short-term NSAID use, and the rate of events is linear over time with continued use. Although gastroprotective therapies are available, they are underused, and patient and physician awareness and recognition of some of the factors influencing the development of NSAID-related upper GI complications are limited. Herein, we present a case report of a patient experiencing a gastric ulcer following NSAID use and examine some of the risk factors and potential strategies for prevention of upper GI mucosal injuries and associated bleeding following NSAID use. These risk factors include advanced age, previous history of GI injury, and concurrent use of medications such as anticoagulants, aspirin, corticosteroids, and selective serotonin reuptake inhibitors. Strategies for prevention of GI injuries include anti-secretory agents, gastroprotective agents, alternative NSAID formulations, and nonpharmacologic therapies. Greater awareness of the risk factors and potential therapies for GI complications resulting from NSAID use could help improve outcomes for patients requiring NSAID treatment.


The role of pitch and temporal diversity in the perception and production of musical sequences.
In two experiments we explored how the dimensions of pitch and time contribute to the perception and production of musical sequences. We tested how dimensional diversity (the number of unique categories in each dimension) affects how pitch and time combine. In Experiment 1, 18 musically trained participants rated the complexity of sequences varying only in their diversity in pitch or time; a separate group of 18 pianists reproduced these sequences after listening to them without practice. Overall, sequences with more diversity were perceived as more complex, but pitch diversity influenced ratings more strongly than temporal diversity. Further, although participants perceived sequences with high levels of pitch diversity as more complex, errors were more common in the sequences with higher diversity in time. Sequences in Experiment 2 exhibited diversity in both pitch and time; diversity levels were a subset of those tested in Experiment 1. Again diversity affected complexity ratings and errors, but there were no statistical interactions between dimensions. Nonetheless, pitch diversity was the primary factor in determining perceived complexity, and again temporal errors occurred more often than pitch errors. Additionally, diversity in one dimension influenced error rates in the other dimension in that both error types were more frequent relative to Experiment 1. These results suggest that although pitch and time do not interact directly, they are nevertheless not processed in an informationally encapsulated manner. The findings also align with a dimensional salience hypothesis, in which pitch is prioritised in the processing of typical Western musical sequences.


Characterization of new potential anticancer drugs designed to overcome glutathione transferase mediated resistance.
Resistance against anticancer drugs remains a serious obstacle in cancer treatment. Here we used novel strategies to target microsomal glutathione transferase 1 (MGST1) and glutathione transferase pi (GSTP) that are often overexpressed in tumors and confer resistance against a number of cytostatic drugs, including cisplatin and doxorubicin (DOX). By synthetically combining cisplatin with a GST inhibitor, ethacrynic acid, to form ethacraplatin, it was previously shown that cytosolic GST inhibition was improved and that cells became more sensitive to cisplatin. Here we show that ethacraplatin is easily taken up by the cells and can reverse cisplatin resistance in MGST1 overexpressing MCF7 cells. A second and novel strategy to overcome GST mediated resistance involves using GST releasable cytostatic drugs. Here we synthesized two derivatives of DOX, 2,4-dinitrobenzenesulfonyl doxorubicin (DNS-DOX) and 4-mononitrobenzenesulfonyl doxorubicin (MNS-DOX) and showed that they are substrates for MGST1 and GSTP (releasing DOX). MGST1 overexpressing cells are resistant to DOX. The resistance is partially reversed by DNS-DOX. Interestingly, the less reactive MNS-DOX was more cytotoxic to cells overexpressing MGST1 than control cells. It would appear that, by controlling the reactivity of the prodrug, and thereby the DOX release rate, selective toxicity to MGST1 overexpressing cells can be achieved. In the case of V79 cells, DOX resistance proportional to GSTP expression levels was noted. In this case, not only was drug resistance eliminated by DNS-DOX but a striking GSTP-dependent increase in toxicity was observed in the clonogenic assay. In summary, MGST1 and GSTP resistance to cytostatic drugs can be overcome and cytotoxicity can be enhanced in GST overexpressing cells.


CD11b+Ly6G- myeloid cells mediate mechanical inflammatory pain hypersensitivity.
Pain hypersensitivity at the site of inflammation as a result of chronic immune diseases, pathogenic infection, and tissue injury is a common medical condition. However, the specific contributions of the innate and adaptive immune system to the generation of pain during inflammation have not been systematically elucidated. We therefore set out to characterize the cellular and molecular immune response in two widely used preclinical models of inflammatory pain: (i) intraplantar injection of complete Freund's adjuvant (CFA) as a model of adjuvant- and pathogen-based inflammation and (ii) a plantar incisional wound as a model of tissue injury-based inflammation. Our findings reveal differences in temporal patterns of immune cell recruitment and activation states, cytokine production, and pain in these two models, with CFA causing a nonresolving granulomatous inflammatory response whereas tissue incision induced resolving immune and pain responses. These findings highlight the significant differences and potential clinical relevance of the incisional wound model compared with the CFA model. By using various cell-depletion strategies, we find that, whereas lymphocyte antigen 6 complex locus G (Ly)6G(+)CD11b(+) neutrophils and T-cell receptor (TCR) β(+) T cells do not contribute to the development of thermal or mechanical pain hypersensitivity in either model, proliferating CD11b(+)Ly6G(-) myeloid cells were necessary for mechanical hypersensitivity during incisional pain, and, to a lesser extent, CFA-induced inflammation. However, inflammatory (CCR2(+)Ly6C(hi)) monocytes were not responsible for these effects. The finding that a population of proliferating CD11b(+)Ly6G(-) myeloid cells contribute to mechanical inflammatory pain provides a potential cellular target for its treatment in wound inflammation.


Space re-exploration in hemispatial neglect.
Exploration of the space around us is a fundamental part of human behaviour. When it breaks down there is an important opportunity to understand its underlying mechanisms. Here we show that many right-hemisphere patients with left neglect re-explore rightward locations, failing to keep track of them during search. Importantly, such re-exploration occurred despite leftward stimuli being indistinguishable in peripheral vision, so it is unlikely to result from implicit processing of neglected targets. Revisits generally occurred after visits to other targets and are therefore not immediate perseverations. Finally, manipulating the visual salience of found targets altered the degree of neglect, but not revisit rates. Space exploration appears to be modulated both by the ability to keep track of spatial locations and by stimulus salience.


Are vascular abnormalities a primary cause or secondary consequence of hypertension?
This paper reviews the evidence that the resistance vasculature is altered in hypertension and the role that the vasculature may play in the pathogenesis of the disease. Although functional changes (i.e., increased vascular smooth muscle sensitivity) have been found to be associated with some models of hypertension (e.g., spontaneously hypertensive rat), in human essential hypertension it appears that the abnormalities that predominate in the resistance vasculature are structural in nature. These changes result in an increased media/lumen ratio of the more proximal resistance vessels (i.d. 100-300 microns), and the changes are such that they could account for many of the altered hemodynamic characteristics seen in patients with essential hypertension (e.g., increased minimum vascular resistance, increased pressor response). However, evidence that the abnormal structure of the peripheral vasculature is a prime determinant of blood pressure is still lacking, and much of the available evidence suggests that the altered structure is a secondary adaptation. Nevertheless, the abnormal vascular structure may play an important pathological role concerning the morbid consequences of the hypertensive disease, suggesting that normalization of vascular structure is a desirable aim for antihypertensive treatment. At present it seems that treatment must be continued for long periods, maybe many years, before vascular structure is normalized.


Outer Plexiform Layer Structures Are Not Altered Following AAV-Mediated Gene Transfer in Healthy Rat Retina.
Ocular gene therapy approaches have been developed for a variety of different diseases. In particular, clinical gene therapy trials for RPE65 mutations, X-linked retinoschisis, and choroideremia have been conducted at different centers in recent years, showing that adeno-associated virus (AAV)-mediated gene therapy is safe, but limitations exist as to the therapeutic benefit and long-term duration of the treatment. The technique of vector delivery to retinal cells relies on subretinal injection of the vector solution, causing a transient retinal detachment. Although retinal detachments are known to cause remodeling of retinal neuronal structures as well as significant cell loss, the possible effects of this short-term therapeutic retinal detachment on retinal structure and circuitry have not yet been studied in detail. In this study, retinal morphology and apoptotic status were examined in healthy rat retinas following AAV-mediated gene transfer via subretinal injection with AAV2/5.CMV.d2GFP or sham injection with fluorescein. Outer plexiform layer (OPL) morphology was assessed by immunohistochemical labeling, laser scanning confocal microscopy, and electron microscopy. The number of synaptic contacts in the OPL was quantified after labeling with structural markers. To assess the apoptotic status, inflammatory and pro-apoptotic markers were tested and TUNEL assay for the detection of apoptotic nuclei was performed. Pre- and postsynaptic structures in the OPL, such as synaptic ribbons or horizontal and bipolar cell processes, did not differ in size or shape in injected versus non-injected areas and control retinas. Absolute numbers of synaptic ribbons were not altered. No signs of relevant gliosis were detected. TUNEL labeling of retinal cells did not vary between injected and non-injected areas, and apoptosis-inducing factor was not delocalized to the nucleus in transduced areas. The neuronal circuits in the OPL of healthy rat retinas undergoing AAV-mediated gene transfer were not altered by the temporary retinal detachment caused by subretinal injection, the presence of viral particles, or the expression of green fluorescent protein as a transgene. This observation likely requires further investigations in the dog model for RPE65 deficiency in order to determine the impact of RPE65 transgene expression on diseased retinas in animals and men.


Affected upper-extremity movement ability is retained 3 months after modified constraint-induced therapy.
The practicality and patient compliance of constraint-induced movement therapy limit its application in many clinical environments. For more than a decade, the principal investigator's laboratory has shown efficacy of an outpatient, modified constraint-induced therapy (mCIT). The current study examined whether participants administered mCIT retained motor changes 3 mo after intervention. The upper-extremity section of the Fugl-Meyer Impairment Scale (FM) and the Action Research Arm Test (ARA) were administered directly after mCIT intervention. Thirteen patients poststroke were tracked prospectively from directly after intervention concluded to 3 mo after intervention, at which time the FM and ARA were readministered. Three months after intervention, 25 of the 26 scores on the FM and ARA increased between the time after intervention and 3 months after intervention, reflecting continued increases in affected extremity movement ability. It is believed that the continued motor changes were caused by the comparatively larger number of extremity-use opportunities during the 10-wk mCIT intervention period. These opportunities encourage habitual extremity use even after the intervention period has concluded, leading to the changes observed.


The ABC of the BX-C: the bithorax complex explained.
As one of two Drosophila Hox clusters, the bithorax complex (BX-C) is responsible for determining the posterior thorax and each abdominal segment of the fly. Through the dissection of its large cis-regulatory region, biologists have obtained a wealth of knowledge that has informed our understanding of gene expression, chromatin dynamics and gene evolution. This primer attempts to distill and explain our current knowledge about this classic, complex locus.


Renal lysosomes: role in biogenesis of erythropoietin.
The "light" mitochondrial pellet obtained from the kidneys of rats previously treated with Triton WR-1339 and rendered hypoxic was separated into subcellular component fractions by sucrose density gradient centrifugation in a zonal rotor. Selected fractions were pooled, disrupted by osmotic lysis and repeated freeze-thawing, and incubated in the presence and absence of normal rat serum. The incubation mixtures were assayed for erythropoiesis-stimulating activity (erythropoietin). High specific activity was identified only in fractions rich in lysosomes. Biochemical analysis of reference enzymes for the identification of lysosomes and mitochondria, supplemented by electron microscopic examination of the various separated fractions, supports the observed requirement for lysosomal constituents in the formation of erythropoietin by the kidney.


Minimal surface scaffold designs for tissue engineering.
Triply-periodic minimal surfaces are shown to be a more versatile source of biomorphic scaffold designs than currently reported in the tissue engineering literature. A scaffold architecture with sheetlike morphology based on minimal surfaces is discussed, with significant structural and mechanical advantages over conventional designs. These sheet solids are porous solids obtained by inflation of cubic minimal surfaces to sheets of finite thickness, as opposed to the conventional network solids where the minimal surface forms the solid/void interface. Using a finite-element approach, the mechanical stiffness of sheet solids is shown to exceed that of conventional network solids for a wide range of volume fractions and material parameters. We further discuss structure-property relationships for mechanical properties useful for custom-designed fabrication by rapid prototyping. Transport properties of the scaffolds are analyzed using Lattice-Boltzmann computations of the fluid permeability. The large number of different minimal surfaces, each of which can be realized as sheet or network solids and at different volume fractions, provides design flexibility essential for the optimization of competing design targets.


HIV-1 Tat-shortened neurite outgrowth through regulation of microRNA-132 and its target gene expression.
Synaptodendritic damage is a pathological hallmark of HIV-associated neurocognitive disorders, and HIV-1 Tat protein is known to cause such injury in the central nervous system. In this study, we aimed to determine the molecular mechanisms of Tat-induced neurite shortening, specifically the roles of miR-132, an important regulator of neurite morphogenesis in this process. The relationship between Tat expression and miR-132 expression was first determined using reverse transcription quantitative PCR (qRT-PCR) in Tat-transfected astrocytes and neurons, astrocytes from Tat-transgenic mice, and HIV-infected astrocytes. qRT-PCR and Western blotting were performed to determine Tat effects on expression of miR-132 target genes methyl CpG-binding protein 2, Rho GTPase activator p250GAP, and brain-derived neurotrophic factor. Exosomes were isolated from Tat-expressing astrocytes, and exosomal microRNA (miRNA) uptake into neurons was studied using miRNA labeling and flow cytometry. The lactate dehydrogenase release was used to determine the cytotoxicity, while immunostaining was used to determine neurite lengths and synapse formation. Tat basic domain deletion mutant and miR-132 mimic and inhibitor were used to determine the specificity of the relationship between Tat and miR-132 and its effects on astrocytes and neurons and the underlying mechanisms of Tat-induced miR-132 expression. Tat significantly induced miR-132 expression, ensuing down-regulation of miR-132 target genes in astrocytes and neurons. miR-132 induction was associated with phosphorylation of cAMP response element-binding protein and required the basic domain of Tat. miRNA-132 induction had no effects on astrocyte activation or survival but was involved in the direct neurotoxicity of Tat. miR-132 was present in astrocyte-derived exosomes and was taken up by neurons, causing neurite shortening. Tat-induced miR-132 expression contributes to both direct and astrocyte-mediated Tat neurotoxicity and supports the important roles of miR-132 in controlling neurite outgrowth.


APOE epsilon 4 influences the pathological phenotype of Alzheimer's disease by favouring cerebrovascular over parenchymal accumulation of A beta protein.
The relative amounts of amyloid beta-protein (A beta) in cerebral blood vessels and parenchyma vary considerably amongst patients with Alzheimer's disease (AD). Although several mechanisms have been proposed to explain this variability, the underlying genetic and environmental determinants are still unclear, as are the functional consequences. Polymorphisms in APOE, the gene for apolipoprotein E (ApoE), influence the risk of developing AD and of deposition of A beta within the brain. We examined the relationship between the APOE genotype and the relative extent of accumulation of A beta as plaques within the cerebral parenchyma and in cortical blood vessels in the form of cerebral amyloid angiopathy (CAA), in autopsy brain tissue from 125 AD cases and from 53 elderly, neurologically normal controls of which 19 had CAA without other neuropathological features of AD. In the AD cases, we also assessed whether the severity of CAA was related to the age of onset and duration of dementia, risk factors for atherosclerotic vascular disease, and histologically demonstrable cerebral infarcts or foci of haemorrhage. The APOE genotype was determined by a standard polymerase chain reaction-based method. Paraffin sections of frontal, temporal and parietal lobes were immunolabelled for A beta and the parenchymal A beta load (total A beta minus vessel-associated A beta) was quantified by computer-assisted image analysis. CAA severity was scored for cortical and leptomeningeal vessels. The relevant clinical data were obtained from the database of the South West Brain Bank. In AD, we found the severity of CAA to be strongly associated with the number of epsilon 4 alleles (P < 0.0001) but the parenchymal A beta load to be independent of APOE genotype. Cases with severe CAA had a lower parenchymal A beta load than had those with moderate CAA (P = 0.003). Neither the severity of CAA nor the parenchymal A beta load correlated with age of onset, duration of disease or age at death, and the severity of CAA also did not correlate with the presence of cerebral infarcts or foci of haemorrhage. These findings indicate that possession of the APOE epsilon 4 allele favours vascular over parenchymal accumulation of A beta in AD. This may influence the pathogenesis of neurodegeneration in epsilon 4-associated AD.


Evaluation of the Humphrey FASTPAC threshold program in glaucoma.
The makers of the Humphrey perimeter have developed a rapid thresholding program, FASTPAC, to address the problems of fatigue and poor test performance associated with prolonged perimetry. The performance of FASTPAC was compared with the standard threshold program of the Humphrey visual field analyser (program 24-2) in 44 glaucoma patients. FASTPAC reduced the mean test time by 35% (from 12.6 to 8.2 minutes), owing to a reduction in the number of stimulus presentations. FASTPAC underestimated the mean deviation (MD) (Wilcoxon, p = 0.007) and corrected pattern standard deviation (CPSD) (Wilcoxon, p = 0.005). The sum of the differences between FASTPAC and the standard program was -1.19 (SD 2.37) dB for MD and 0.97 (2.14) dB for CPSD. The measured difference between the two methods was independent of the value of either MD or CPSD, indicating that the measurement error was just as likely to occur with either early or advanced visual field loss. No difference was noted in short term fluctuation or in the reliability indices of test performance. These results indicate that FASTPAC, while considerably faster, is not as accurate as the standard threshold program at measuring retinal sensitivity in glaucoma. This inaccuracy may be offset in practical terms by greater patient acceptability and by increasing the number of patients capable of performing reliable threshold perimetry because of the reduction in duration of the FASTPAC program.


Surface exposure of phosphatidylserine in pathological cells.
The asymmetric phospholipid distribution in plasma membranes is normally maintained by energy-dependent lipid transporters that translocate different phospholipids from one monolayer to the other against their respective concentration gradients. When cells are activated, or enter apoptosis, lipid asymmetry can be perturbed by other lipid transporters (scramblases) that shuttle phospholipids non-specifically between the two monolayers. This exposes phosphatidylserine (PS) at the cells' outer surface. Since PS promotes blood coagulation, defective scramblase activity upon platelet stimulation causes a bleeding disorder (Scott syndrome). PS exposure also plays a pivotal role in the recognition and removal of apoptotic cells via a PS-recognizing receptor on phagocytic cells. Furthermore, expression of PS at the cell surface can occur in a wide variety of disorders. This review aims at highlighting how PS expression in different cells may complicate a variety of pathological conditions, including those that promote thromboembolic complications or produce aberrations in apoptotic cell removal.


Determinants of the geographical distribution of endemic giardiasis in Ontario, Canada: a spatial modelling approach.
Giardiasis surveillance data as well as drinking water, socioeconomic and land-use data were used in spatial regression models to investigate determinants of the geographic distribution of endemic giardiasis in southern Ontario. Higher giardiasis rates were observed in areas using surface water [rate ratio (RR) 2.36, 95 % CI 1.38-4.05] and in rural areas (RR 1.79, 95 % CI 1.32-2.37). Lower rates were observed in areas using filtered water (RR 0.55, 95 % CI 0.42-0.94) and in those with high median income (RR 0.62, 95 % CI 0.42-0.92). Chlorination of drinking water, cattle density and intensity of manure application on farmland were not significant determinants. The study shows that waterborne transmission plays an important role in giardiasis distribution in southern Ontario and that well-collected routine surveillance data could be useful for investigation of disease determinants and identification of high-risk communities. This information is useful in guiding decisions on control strategies.


Beyond fragmentation and towards universal coverage: insights from Ghana, South Africa and the United Republic of Tanzania.
The World Health Assembly of 2005 called for all health systems to move towards universal coverage, defined as " access to adequate health care for all at an affordable price" . A crucial aspect in achieving universal coverage is the extent to which there are income and risk cross-subsidies in health systems. Yet this aspect appears to be ignored in many of the policy prescriptions directed at low- and middle-income countries, often resulting in high degrees of health system fragmentation. The aim of this paper is to explore the extent of fragmentation within the health systems of three African countries (Ghana, South Africa and the United Republic of Tanzania). Using a framework for analysing health-care financing in terms of its key functions, we describe how fragmentation has developed, how each country has attempted to address the arising equity challenges and what remains to be done to promote universal coverage. The analysis suggests that South Africa has made the least progress in addressing fragmentation, while Ghana appears to be pursuing a universal coverage policy in a more coherent way. To achieve universal coverage, health systems must reduce their reliance on out-of-pocket payments, maximize the size of risk pools, and resource allocation mechanisms must be put in place to either equalize risks between individual insurance schemes or equitably allocate general tax (and donor) funds. Ultimately, there needs to be greater integration of financing mechanisms to promote universal cover with strong income and risk cross-subsidies in the overall health system.


Methylglyoxal inhibits seed germination and root elongation and up-regulates transcription of stress-responsive genes in ABA-dependent pathway in Arabidopsis.
Methylglyoxal (MG) is a highly reactive metabolite derived from glycolysis. In this study, we examined the effect of MG on seed germination, root elongation, chlorosis and stress-responsive gene expression in Arabidopsis using an abscisic acid (ABA)-deficient mutant, aba2-2. In the wild type, 0.1 mm MG did not affect germination but delayed root elongation, whereas 1.0 mm MG inhibited germination and root elongation and induced chlorosis. MG increased transcription levels of RD29B and RAB18 in a dose-dependent manner but did not affect RD29A transcription level. In contrast, in the aba2-2 mutant, MG inhibition of seed germination at 1.0 mm and 10.0 mm and a delay of root elongation at 0.1 mm MG were mitigated, although there was no significant difference in chlorosis between the wild type and mutant. Moreover, the aba2-2 mutation impaired MG-induced RD29B and RAB18 gene expression. These observations suggest that MG not only directly inhibits germination and root elongation but also indirectly modulates these processes via endogenous ABA in Arabidopsis.


Synthesis of 1L-chiro-inositol 2,3,5-trisphosphorothioate, the first partial agonist at the platelet myo-inositol 1,4,5-trisphosphate receptor.
The synthesis of L-chiro-inositol 2,3,5-trisphosphorothioate, a novel analogue of the second messenger D-myo-inositol 1,4,5-trisphosphate has been accomplished from the natural product L-quebrachitol. Phosphitylation of (-)-1L-1,4,6-tri-O-benzoyl-chiro-inositol obtained from L-quebrachitol followed by sulfoxidation of the products gave (-)-1L-1,4,6-tri-O-benzoyl-chiro-inositol 2,3,5-tris[di(2-cyanoethyl) phosphorothioate], which was deblocked using sodium in liquid ammonia to give 1L-(-)-chiro-inositol 2,3,5-trisphosphorothioate. 1L-chiro-Inositol 2,3,5-trisphosphorothioate is a partial agonist in the release of intracellular Ca2+ from saponin-permeabilised platelets and is both a key tool for pharmacological dissection of the polyphosphoinositide pathway of cellular signalling and a lead compound for the design of small molecule Ins(1,4,5)P3 receptor antagonists.


[Effect of phlebodium decumanum and coenzyme Q10 on sports performance in professional volleyball players].
Physical training programmes are based on provoking transitory states of fatigue in order to induce super compensation by the biological systems involved in the activity, in order to improve the athlete's medium-long term performance. The administration of nutritional supplements with antioxidant and immunomodulatory properties, such as Phlebodium decumanum and coenzyme Q10, can be a very advantageous means of achieving recovery from the inflammation and tissue damage caused by the stress of prolonged, intense exercise. An experimental, longitudinal, double- blind experiment was conducted, with three randomised groups obtained from a sample of 30 male volleyball players (aged 22-32 years) at the University of Granada, with a high level of training (17 hours a week during the 6 months preceding the study). The effects were then evaluated of a month-long physical training programme, common to all the study groups, associated with the simultaneous administration of the following nutritional supplements: Phlebodium decumanum (4 capsules of 400 mg/capsule, daily), Experimental Group 1; Phlebodium decumanum (same dose and schedule as Group 1) plus coenzyme Q10 (4 capsules of 30 mg/ capsule, daily), Experimental Group 2; a placebo substance, Control Group. The following dependent blood variables were examined to assess the effects of the intervention on the basal immune and endocrine-metabolic profile: cortisol and interleukin-6, both related to the axis of exercise-induced stress; and lactic acid and ammonium, related essentially to the anaerobic metabolism of energy. All the study groups presented favourable adaptive changes with respect to the endocrine-metabolic and immune profile, as reflected by a significant decrease in the post-test concentrations of cortisol, interleukin 6, lactic acid and ammonium, compared to the values recorded before the physical activity with/without nutritional supplement, per protocol. The groups that achieved the most favourable profile were those which had received nutritional supplementation, rather than the placebo, and among the former, those which had received the double- strength supplement with Phlebodium decumanum plus coenzyme Q10. The intake of Phlebodium decumanum plus coenzyme Q10 for 4 weeks produced protective effects on the endocrine-metabolic and immune profile, which we attribute to the immunomodulatory and antioxidant properties of these substances, which are highly beneficial not only in terms of delaying fatigue and improving athletic performance, but also in reducing the risk of injuries associated with high intensity exercise.


Mass spectrometric techniques for the analysis of volatile organic compounds emitted from bacteria.
Bacteria are the main cause of many human diseases. Typical bacterial identification methods, for example culture-based, serological and genetic methods, are time-consuming, delaying the potential for an early and accurate diagnosis and the appropriate subsequent treatment. Nevertheless, there is a stringent need for in situ tests that are rapid, noninvasive and sensitive, which will greatly facilitate timely treatment of the patients. This review article presents volatile organic metabolites emitted from various micro-organism strains responsible for common bacterial infections in humans. Additionally, the manuscript shows the application of different analytical techniques for fast bacterial identification. Details of these techniques are given, which focuses on their advantages and drawbacks in using for volatile organic components analysis.


The Effects of Aging on Corneal and Ocular Surface Homeostasis in Mice.
Aging is a risk factor for dry eye disease. The aim of this study was to investigate if aging is associated with a range of signs of dry eye disease, including tear hyperosmolarity, reduced nerve density, and increased dendritic cell number, in mice. Healthy C57BL/6 female mice, aged 2 months (young, n = 10) and 22 months (aged, n = 11) were used. Clinical assessments included corneal sensitivity (Cochet-Bonnet esthesiometry), tear secretion (Phenol red thread test), tear film osmolarity (TearLab osmometer), and corneal thickness (optical coherence tomography). The sum length of the corneal superficial terminals and sub-basal nerves, density of vertical nerve projections, and density and tree area of resident epithelial dendritic cells, were assessed using immunofluorescence and confocal microscopy. Aged mice had significantly higher tear secretion, lower corneal sensitivity, and a thicker corneal stroma but thinner epithelium. There was no significant intergroup difference for tear osmolarity. Aged mice showed a significantly lower sum length of nerves in the superficial terminals and sub-basal plexus, relative to young mice. Dendritic cell density and morphology were similar in both groups. In mice, aging is associated with higher tear secretion and corneal epithelial thinning, together with lower corneal nerve density and sensitivity. However, aging was not significantly associated with changes to tear osmolarity or dendritic cell density or size, despite a significant reduction in total nerve length. These findings demonstrate that aged mice exhibit some changes to ocular surface parameters that parallel the anomalies evident in dry eye disease.


Differences in menopausal hormone therapy use among women in Germany between 1998 and 2003.
To examine the differences in menopausal hormone therapy (MHT) use and user profiles among women in Germany before and after the communication of the Women's Health Initiative (WHI) trial and other study results concerning the risks and benefits of MHT. Current MHT use was ascertained in two periodic German national health surveys conducted in 1997-1999 and 2003-2004. MHT prevalence and user profiles were assessed within each survey. The association of the survey period (2003-2004 vs. 1997-1999) with current MHT use was analyzed in weighted multivariable logistic regression (MLR) models, pooling data from both surveys. The overall prevalence of current MHT use decreased by 40.2% from 16.9% of the sample in 1997-1999 to 10.1% in 2003-2004. The difference in prevalence between surveys varied with age decade with the smallest decreases among women 60-69 years of age (20.3% vs. 18.5%), compared to women of younger and older age groups (40-49: 10.7% vs. 3.9%; 50-59: 36.3% vs. 21.3%; 70-79: 5.7% vs. 3.2%). Variables independently associated with higher current MHT use in both health surveys included age category (curvilinear relationship with highest use among women 50-59 years) and residence in West vs. East Germany. A higher social status, lower body mass index, and more health-conscious behaviour were significantly associated with higher current MHT use in the 1997-1999 survey, but these associations were not found in the later survey. MLR analyses confirmed a significant decline in MHT use between the 1997-1999 and 2003-2004 surveys, however, the effect was modified by social status and was not significant among lowest social-status women. Current MHT use considerably declined among women in Germany between the pre- and post-WHI era. A convergence of current MHT use among women of higher social status with pre-existing patterns of use among lower social-status women suggests that MHT in Germany is now less likely to be used for health promotion.


Current Practice of Surgery for Benign Goitre-An Analysis of the Prospective DGAV StuDoQ|Thyroid Registry.
To evaluate the current indications, resection strategies and short-term outcomes of surgery for benign goitre in a country with endemic goitre. Data of patients who underwent surgery for benign goitre were retrieved from the prospective StuDoQ/Thyroid registry and retrospectively analysed regarding the patient's demographics, indications for surgery, surgical procedures, histology, and perioperative outcomes. In a 15-month period, 12,888 patients from 83 departments underwent thyroid resections for benign conditions. Main indications for surgery were exclusion of malignancy (68%), compression symptoms (20.7%) and hyperthyroidism (9.7%). Preoperative fine needle aspiration cytology was performed in only 12.2% of patients with the indication "exclusion of malignancy". Thyroidectomy (49.8%) or hemithyroidectomy (36.9%) were performed in 86.7% of patients. Minimally invasive or alternative surgical techniques were applied in only 2.2%. Intraoperative neuromonitoring was used in 98.4% of procedures, in 97.5% of patients at least one parathyroid gland was visualized, and in 15.3% of patients parathyroid tissue was autografted, respectively. The rates of unilateral and bilateral transient recurrent nerve palsy were 3.6% and 0.07% of nerves at risk, the rate of transitory hypoparathyroidism was 15.3%. The rates of postoperative bleeding and wound infections requiring reoperation were 1.4% and 0.07%, respectively. The indication "exclusion of malignancy" is made too liberally, and there is a strong attitude to perform complete thyroid resections. Postoperative hypoparathyroidism is the major complication after surgery for benign thyroid disease, thus requiring more awareness.


Survival in Out-of-hospital Rapid Sequence Intubation of Non-Traumatic Brain Pathologies.
Rapid sequence intubation (RSI) is not only used in traumatic brain injuries in the out-of-hospital setting, but also for non-traumatic brain pathologies (NTBP) such as brain tumors, meningitis, encephalitis, hypoxic/anoxic brain injury, stroke, arteriovenous malformations, tumors, aneurysms, brain hemorrhage, as well as brain injury due to diabetes, seizures and toxicity, metabolic conditions, and alcohol and drug overdose. Previous research suggests that RSI is common in non-traumatic coma, but with an unknown prevalence of NTBP in those that receive RSI. If NTBP is common and if brain trauma RSI evidence is not valid for NTBP then a sizable proportion of NTBP receive this treatment without evidence of benefit. This study calculated the out-of-hospital NTBP prevalence in patients that had received RSI and explored factors that predicted survival. A retrospective cohort study based on data collected from an ambulance service and seven hospitals based in Melbourne, Australia. Non-traumatic brain pathologies were defined using ICD10-AM codes for the calculation of NTBP prevalence. Logistic regression modelled out-of-hospital predictors of survival to hospital discharge after adjustment for comorbidities. The seven participating hospitals treated 2,277 patients that received paramedic RSI for all illnesses and indications from January 1, 2008 to December 31, 2015, with survival data available for 1,940 (85%). Of the 1,940, 1,125 (58%) patients had at least one hospital-diagnosed NTBP. Sixty-nine percent all of NTBP survived to hospital discharge, compared to 65% for traumatic intracranial injury. Strokes were the most common and had poor survival to discharge (37%) compared to the second most common NTBP toxicity/toxic encephalopathy that had very high survival (98%). No out-of-hospital clinical intervention or prehospital time interval predicted survival. Factors that did predict survival include Glasgow Coma Scale (GCS), duration of mechanical ventilation, age, ICU length of stay, and comorbidities. Non-traumatic brain pathologies are seven times more prevalent than traumatic brain injuries in patients that underwent out-of-hospital RSI in Victoria, Australia. Since the mechanisms through which RSI impacts mortality might differ between traumatic brain injuries and NTBP, and given that NTBP is very prevalent, it follows that the use of RSI in NTBP could be unsupported.


Two Pif1 Family DNA Helicases Cooperate in Centromere Replication and Segregation in Saccharomyces cerevisiae.
Pif1 family helicases are found in virtually all eukaryotes. Saccharomyces cerevisiae (Sc) encodes two Pif1 family helicases, ScPif1 and Rrm3 ScPif1 is multifunctional, required not only for maintenance of mitochondrial DNA but also for multiple distinct nuclear functions. Rrm3 moves with the replication fork and promotes movement of the fork through ∼1400 hard-to-replicate sites, including centromeres. Here we show that ScPif1, like Rrm3, bound robustly to yeast centromeres but only if the centromere was active. While Rrm3 binding to centromeres occurred in early to mid S phase, about the same time as centromere replication, ScPif1 binding occurred later in the cell cycle when replication of most centromeres is complete. However, the timing of Rrm3 and ScPif1 centromere binding was altered by the absence of the other helicase, such that Rrm3 centromere binding occurred later in pif1-m2 cells and ScPif1 centromere binding occurred earlier in rrm3Δ cells. As shown previously, the modest pausing of replication forks at centromeres seen in wild-type cells was increased in the absence of Rrm3 While a lack of ScPif1 did not result in increased fork pausing at centromeres, pausing was even higher in rrm3Δ pif1Δ cells than in rrm3Δ cells. Likewise, centromere function as monitored by the loss rate of a centromere plasmid was increased in rrm3Δ but not pif1Δ cells, and was even higher in rrm3Δ pif1Δ cells than in rrm3Δ cells. Thus, ScPif1 promotes centromere replication and segregation, but only in the absence of Rrm3 These data also hint at a potential post-S phase function for ScPif1 at centromeres. These studies add to the growing list of ScPif1 functions that promote chromosome stability.


Associations between Physical Activity and Submaximal Cardiorespiratory and Pulmonary Responses in Men.
BACKGROUND: Habitual physical activity (PA) is associated with higher cardiorespiratory fitness values, but additional information is needed on the contributions of specific types and amounts of PA. Therefore the main aim of this study was to analyze the heart and lung function of a large cohort of men and compare these outcomes with various modes and volumes of PA. METHODS: We used data from 30,594 men from the Aerobics Center Longitudinal Study who were categorized into sedentary, swimmer, walker, and runner groups using self-report PA data collected during 1970-2005. Additional PA categories using MET-minutes/week were used to group men into 5 distinct levels of activity (0 MET-min, 1-499 MET-min, 500-999 MET-min, 1000-1499 MET-min, and ≥ 1500 MET-min). Each participant also completed a maximal treadmill exercise test to quantify their fitness level. Cross-sectional analyses included general linear modeling and multiple comparisons adjusted for age, smoking status, and histories of myocardial infarction, stroke, diabetes, and hypertension. RESULTS: A dose-response linear effect was found for heart function variables across PA MET-min categories. Stronger associations for resting heart rate (HR), heart rate recovery (HRR), exercise HR, and exercise blood pressure were found with the runner and swimming groups when compared to the walkers and sedentary groups. Walkers had significantly better heart function than the sedentary group but only about half the effect seen in the swimmers and runners. Lung function findings showed greater absolute values in FVC and FEV1 across PA categories, but found no difference in lung function ratios (e.g FEV1/FVC%). CONCLUSIONS: We found beneficial linear associations with resting HR, exercise HR, HRR, fitness values, FVC, and FEV1 over increasing MET-min categories. This implies that habitual PA, such as walking, but especially swimming and running, when performed with adequate volume, are viable ways to gain benefits for heart health.


Studies on the cross-striation of the indirect flight myofibrils of the blowfly Calliphora.
1. The cross-striation in the indirect flight myofibrils of Calliphora has been studied by phase contrast and polarised light microscopy. The band pattern at rest-length has been determined in flies killed in osmium tetroxide vapour while their wings remained in the resting position. All other observations have been made on unfixed fibrils. Although length changes in situ are probably very slight (about 2 per cent), isolated fibrils, by treatment with crude muscle extract or with ATP, can be induced to elongate to 104 per cent rest-length, or to shorten by 8 per cent but no more. Over the range 98 to 104 per cent rest-length, experimentally induced length changes are reversible. The fibrils can also be stretched beyond 104 per cent rest-length, but the process is irreversible. During the course of glycerol extraction the fibrils elongate to 104 per cent rest-length. 2. The changes in band pattern observed over the range 104 to 92 per cent rest-length are qualitatively the same as the changes observed over a wider range (about 130 to 40 per cent rest-length) in the skeletal myofibrils of rabbits. The earlier stages of shortening appear to be effected by retraction of the I bands into the A bands where they fill up the H zones. No evidence has been found that any changes in band pattern are due to a migration of the A substance. 3. Two components of the sarcomere can be extracted from it and a third component remains behind. These three components, which have also been demonstrated in skeletal myofibrils of the rabbit, where they behave in the same way, are: (a) the A substance which does not change its position as the fibril changes its length, and which can be extracted by the same procedures as remove myosin (shown elsewhere to be the A substance) from rabbit fibrils; (b) a material which extends from the Z lines to the borders of the H zone and which moves inwards during contraction and outwards during elongation; it can capture rabbit myosin from solution and form with it a contractile system, and it is thought to be actin; (c) a "backbone" or stroma bearing Z and M lines. 4. Since all these features of the cross-striation are the same in the insect fibrils as in rabbit fibrils, it is considered very probable that the sarcomere is similarly organised in both types of muscle and contracts by essentially the same mechanism.


Characterization of the bacteriophage phi29-encoded protein p16.7: a membrane protein involved in phage DNA replication.
An early expressed operon, located at the right end of the linear bacteriophage phi29 genome, contains open reading frame (ORF)16.7, whose deduced protein sequence of 130 amino acids is conserved in phi29-related phages. Here, we show that this ORF actually encodes a protein, p16.7, which is abundantly and early expressed after infection. p16.7 is a membrane protein, and the N-terminally located transmembrane-spanning domain is required for its membrane localization. The variant p16.7A, in which the N-terminal membrane anchor was replaced by a histidine-tag, was purified and characterized. Purified p16.7A was shown to form dimers in solution. To study the in vivo role of p16.7, a phi29 mutant containing a suppressible mutation in gene 16.7 was constructed. In vivo phage DNA replication was affected in the absence of p16.7, especially at early infection times. Based on the results, the putative role of p16.7 in in vivo phi29 DNA replication is discussed.


Health-care facility choice and the phenomenon of bypassing.
Health policy-makers in developing countries are often disturbed and to a degree surprised by the phenomenon of the ill travelling past a free or subsidized local public clinic (or other public facility) to get to an alternative source of care at which they often pay a considerable amount for health care. That a person bypasses a facility is almost certainly indicative either of significant problems with the quality of care at the bypassed facility or of significantly better care at the alternative source of care chosen. When it is a poor person choosing to bypass a free public facility and pay for care further away, such action is especially bothersome to public policy-makers. This paper uses a unique data set, with a health facility survey in which all health facilities are identified, surveyed, and located geographically; and a household survey in which a sample of households from the same health district is also both surveyed and located geographically. The data are analyzed to examine patterns of health care choice related to the characteristics and locations of both the facilities and actual and potential clients. Rather than using the distance travelled or some other general choice of type of care variable as the dependent variable, we are able actually to analyze which specific facilities are bypassed and which chosen. The findings are instructive. That bypassing behaviour is not very different across income groups is certainly noteworthy, as is the fact that the more severely ill tend to bypass and to travel further for care than do the less severely ill. In multivariate analysis almost all characteristics of both providers and facilities are found to have the a priori expected relationships to facility choice. Prices tend to deter use, and improved quality of services to increase the likelihood of a facility being chosen. The answer to the bypassing dilemma seems to be for providers to provide as good quality care relative to the money charged (if any), as other, often further away, providers.


Hepatic safety of maraviroc in patients with HIV-1 and hepatitis C and/or B virus: 144-week results from a randomized, placebo-controlled trial.
In the primary 48-week analysis of a hepatic safety trial in patients with HIV-1 coinfected with HBV and/or HCV, maraviroc-containing treatment regimens were not associated with increased hepatotoxicity. In this randomized, double-blind, placebo-controlled, multicentre study, patients received maraviroc twice daily (n=70) or placebo (n=67) with concomitant antiretroviral therapy for 144 weeks (Clinicaltrials.gov identifier, NCT01327547). The primary end point was the proportion of patients with protocol-defined Grade 3/4 alanine aminotransferase (ALT) abnormalities through week 48. Key secondary end points included 144-week analysis of Grade 3/4 ALT abnormalities and liver fibrosis by enhanced liver fibrosis (ELF) testing, hepatic elastography and an optional biopsy substudy. Through 144 weeks of treatment, two (maraviroc) and three (placebo) patients met the protocol-defined Grade 3/4 ALT end point. Similar to the 48-week results, there were no statistically significant differences between groups in change from baseline in ELF or hepatic elastography. However, decreased elastography scores were noted in the maraviroc group. Blinded pathologist review suggested that 2 of 11 paired biopsies (both on maraviroc) showed signs of decreased fibrosis. One (maraviroc) and two (placebo) patients experienced treatment-related hepatobiliary adverse events (AEs). Five patients in the maraviroc group discontinued because of treatment-related AEs versus three in the placebo group. One death in the maraviroc group and two deaths in the placebo group were reported. Use of maraviroc did not increase hepatotoxicity in this population through 144 weeks. Further investigation regarding possible beneficial effects of maraviroc on liver fibrosis may be warranted.


FMNL2 and -3 regulate Golgi architecture and anterograde transport downstream of Cdc42.
The Rho-family small GTPase Cdc42 localizes at plasma membrane and Golgi complex and aside from protrusion and migration operates in vesicle trafficking, endo- and exocytosis as well as establishment and/or maintenance of cell polarity. The formin family members FMNL2 and -3 are actin assembly factors established to regulate cell edge protrusion during migration and invasion. Here we report these formins to additionally accumulate and function at the Golgi apparatus. As opposed to lamellipodia, Golgi targeting of these proteins required both their N-terminal myristoylation and the interaction with Cdc42. Moreover, Golgi association of FMNL2 or -3 induced a phalloidin-detectable actin meshwork around the Golgi. Importantly, functional interference with FMNL2/3 formins by RNAi or CRISPR/Cas9-mediated gene deletion invariably induced Golgi fragmentation in different cell lines. Furthermore, absence of these proteins led to enlargement of endosomes as well as defective maturation and/or sorting into late endosomes and lysosomes. In line with Cdc42 - recently established to regulate anterograde transport through the Golgi by cargo sorting and carrier formation - FMNL2/3 depletion also affected anterograde trafficking of VSV-G from the Golgi to the plasma membrane. Our data thus link FMNL2/3 formins to actin assembly-dependent functions of Cdc42 in anterograde transport through the Golgi apparatus.


Total Flavonoids of Drynariae Rhizoma Prevent Bone Loss Induced by Hindlimb Unloading in Rats.
Drynariae Rhizoma is a kidney-tonifying herb that has a long history in clinical practice for the treatment of bone fractures and joint diseases in China. Flavonoids are considered to be its major active ingredients and are reported to ease bone loss in ovariectomized rats. However, the beneficial effects of the total flavonoids of Drynariae Rhizoma on osteoporosis caused by microgravity or mechanical inactivity remain unknown. This study assessed the effects of total Drynariae Rhizoma flavonoids (DRTF, Qihuang, Beijing, China, national medicine permit No. Z20030007, number of production: 04080081, content of DRTF ≥80%) against bone loss induced by simulated microgravity. A hindlimb unloading tail-suspended rat model was established to determine the effect of DRTF on bone mineral density (BMD), biomechanical strength and trabecular bone microarchitecture. Twenty-eight male Sprague-Dawley rats were divided into four groups: the baseline, control, hindlimb unloading with vehicle (HLU), and hindlimb unloading treated with DRTF (HLU-DRTF, 75 mg/kg/day) groups. Oral DRTF was administered for 4 weeks. The underlying mechanisms of the DRTF actions on disuse-induced osteoporosis are discussed. The results showed that DRTF treatment significantly increased the BMD and mechanical strength of tail-suspended rats. Enhanced bone turnover markers with HLU treatment were attenuated by DRTF administration. Deterioration of trabecular bone induced by HLU was prevented through elevated bone volume/tissue volume (BV/TV), trabecular number (Tb. N), trabecular thickness (Tb. Th) and decreased trabecular separation (Tb. Sp). The present study provides the first evidence that DRTF prevents bone loss induced by HLU treatment, indicating its potential application in the treatment of disuse-induced osteoporosis.


Deep white matter analysis: fast, consistent tractography segmentation across populations and dMRI acquisitions.
We present a deep learning tractography segmentation method that allows fast and consistent white matter fiber tract identification across healthy and disease populations and across multiple diffusion MRI (dMRI) acquisitions. We create a large-scale training tractography dataset of 1 million labeled fiber samples (54 anatomical tracts are included). To discriminate between fibers from different tracts, we propose a novel 2D multi-channel feature descriptor (FiberMap) that encodes spatial coordinates of points along each fiber. We learn a CNN tract classification model based on FiberMap and obtain a high tract classification accuracy of 90.99%. The method is evaluated on a test dataset of 374 dMRI scans from three independently acquired populations across health conditions (healthy control, neuropsychiatric disorders, and brain tumor patients). We perform comparisons with two state-of-the-art white matter tract segmentation methods. Experimental results show that our method obtains a highly consistent segmentation result, where over 99% of the fiber tracts are successfully detected across all subjects under study, most importantly, including patients with space occupying brain tumors. The proposed method leverages deep learning techniques and provides a much faster and more efficient tool for large data analysis than methods using traditional machine learning techniques.


Distinct Trauma Types in Military Service Members Seeking Treatment for Posttraumatic Stress Disorder.
We examined the frequency of trauma types reported in a cohort of service members seeking treatment for posttraumatic stress disorder (PTSD) and compared symptom profiles between types. In this observational study, 999 service members (9.2% women; Mage = 32.91 years; 55.6% White) were evaluated using a standardized assessment procedure to determine eligibility for clinical trials. Participants were evaluated for DSM-IV-TR-defined PTSD using the PTSD Symptom Scale-Interview; all participants reported a Criterion A event. Independent evaluators rated descriptions of Criterion A events as belonging to trauma types at a high degree of reliability, κ = 0.80. Aggregated non-life-threat primary trauma types were more frequently endorsed than aggregated life-threat types, 95% CI [17.10%, 29.20%]. Participants who endorsed moral injury-self traumas had a higher level of reexperiencing (d = 0.39), guilt (hindsight bias, d = 1.06; wrongdoing, d = 0.93), and self-blame (d = 0.58) symptoms, relative to those who reported life threat-self. Participants who experienced traumatic loss had greater reexperiencing (d = 0.39), avoidance (d = 0.22), guilt (responsibility, d = 0.39), and greater peri- and posttraumatic sadness (d = 0.84 and d = 0.70, respectively) symptoms, relative to those who endorsed life threat-self. Relative to life threat-self, moral injury-others was associated with greater peri- (d = 0.36) and posttraumatic (d = 0.33) betrayal/humiliation symptoms, and endorsement of aftermath of violence was associated with greater peri- (d = 0.84) and posttraumatic sadness (d = 0.57) symptoms. War zone traumas were heterogeneous, and non-life-threat traumas were associated with distinct symptoms and problems.