NSF is required for transport from early to late endosomes.
Protein transport between early and late endosomes is a major membrane trafficking pathway in the cell followed by many proteins, including all down-regulated receptors. Yet, little is known at the molecular level about the mechanisms regulating membrane interactions in the endocytic pathway beyond early endosomes. In this study, we have used an in vitro transport assay to study the biochemical properties of endosome docking/fusion events. Our data demonstrate that N-ethylmaleimide (NEM) sensitive factor (NSF) and its soluble associated proteins (SNAPs) are required for transport from early to late endosomes, as well as at all other steps of endosomal membrane transport. We also find that these proteins are enriched on endosomal membranes. In addition, our studies suggest that besides NSF/SNAPs, another NEM-sensitive component may also be involved in docking/fusion at this late stage of the pathway. Finally, we find that, in contrast to Golgi membranes, NSF association to both early and late endosomal membranes occurs via an ATP-independent mechanism, indicating that the binding properties of endosomal and biosynthetic NSF are different. Our data thus show that NSF/SNAPs, perhaps together with another NEM-sensitive factor, are part of the basic molecular machinery which controls docking/fusion events during transport from early to late endosomes, along the lysosomal degradation pathway.


Greenhouse gas emissions and global warming potential of reclaimed forest and grassland soils.
Although greenhouse gas (GHG) emissions from soils are important, reclaimed mine soil (RMS) ecosystems are not widely assessed. Postreclamation land uses (forest, hay, and pasture) were investigated to: (i) monitor the magnitude of GHG fluxes, (ii) estimate their global warming potential (GWP), (iii) identify the relationship between GHG fluxes and soil properties, and (iv) develop a soil quality index by principal component analysis (PCA). The GHG fluxes were measured for 1 yr cycle and simultaneous measurements were also made for soil moisture and temperature. The RMS-forest, -hay, and -pasture land uses had weighted average fluxes of 1.16, 1.66, and 3.06 g CO(2)-C m(-2) d(-1); 0.33, 0.48 and 1.1 mg CH(4)-C m(-2) d(-1); and 0.33, 0.70, and 1.06 mg N(2)O-N m(-2) d(-1), respectively. The CO(2), CH(4), and N(2)O fluxes were consistently high in the RMS-pasture and low in the RMS-forest. The GWP (CO(2)-C equivalent) of the postreclamation land uses was in the order of RMS-forest (4.5 Mg ha(-1) yr(-1)) = RMS-hay (6.8 Mg ha(-1) yr(-1)) < RMS-pasture (12.3 Mg ha(-1) yr(-1)). The PCA showed that four PCs with eigenvalues > 1 explained 88.8% of the total variance in the soil properties. The first PC is mostly characterized by soil physical properties and the second by chemical properties. Soil and air temperatures were positively correlated with CO(2), CH(4), and N(2)O fluxes. The results suggest that GWP from RMS can be minimized by establishing forest land use.


A nodule-specific protein secretory pathway required for nitrogen-fixing symbiosis.
The nitrogen-fixing symbiosis between Sinorhizobium meliloti and its leguminous host plant Medicago truncatula occurs in a specialized root organ called the nodule. Bacteria that are released into plant cells are surrounded by a unique plant membrane compartment termed a symbiosome. We found that in the symbiosis-defective dnf1 mutant of M. truncatula, bacteroid and symbiosome development are blocked. We identified the DNF1 gene as encoding a subunit of a signal peptidase complex that is highly expressed in nodules. By analyzing data from whole-genome expression analysis, we propose that correct symbiosome development in M. truncatula requires the orderly secretion of protein constituents through coordinated up-regulation of a nodule-specific pathway exemplified by DNF1.


Family origin and mortality: prospective Finnish cohort study.
Death rates are notably higher in eastern Finland than in western Finland, and life expectancy of Finnish speakers shorter than that of Swedish speakers. The mortality differences correspond to recent genetic mappings of the population and are prominent for causes of death that are known to be associated with genetic risk factors. Using intergenerational data, we studied the impact of parental birth area on all-cause mortality risks of middle-aged men in Finland 1985-2003, assuming that geographic family origin reflects genetic predisposition to complex disorders. Relative death risks at ages 30-49 years were estimated by parental birth region and ethnicity, according to Cox regressions standardised for own education, family type at childhood, and year of birth. The death risk of Finnish speakers born in eastern Finland was 1.13 (95% confidence interval 1.01 to 1.26) that of Finnish speakers born in western Finland, whereas that of Swedish speakers was only 0.60 (0.52 to 0.71). In Finnish speakers, the effects of own birth area and area of residence disappeared when parental birth area was accounted for. The death risk of persons with at least one parent born in eastern Finland was 1.23 (1.09 to 1.39) that of people with both parents born in western Finland. Parental birth area is the driving force behind the regional mortality difference in Finland. The findings highlight and give further support for the potentially important role of genetic risk factors in mortality. Close monitoring of persons' geographic and ethnic ancestry may promote public health and avoid many early deaths.


Acylative Dynamic Kinetic Resolution of Secondary Alcohols: Tandem Catalysis by HyperBTM and Bäckvall's Ruthenium Complex.
Non-enzymatic dynamic kinetic resolution (DKR) of secondary alcohols by enantioselective acylation using an isothiourea-derived HyperBTM catalyst and racemization of slowly reacting alcohol by Bäckvall's ruthenium complex is reported. The DKR approach features high enantioselectivities (up to 99:1), employs easy-to-handle crystalline 4-nitrophenyl isobutyrate as the acylating reagent, and proceeds at room temperature and under an ambient atmosphere. The stereoinduction model featuring cation-π system interactions between the acylated HyperBTM catalyst and π electrons of an alcohol aryl subunit has been elaborated by DFT calculations.


Comparison between modified Dunn procedure and in situ fixation for severe stable slipped capital femoral epiphysis.
Background and purpose - The best treatment option for severe slipped capital femoral epiphysis (SCFE) is still controversial. We compared clinical and radiographic outcomes of modified Dunn procedure (D) and in situ fixation (S) in severe SCFE. Patients and methods - We retrospectively compared D and S, used for severe stable SCFE (posterior sloping angle (PSA) > 50°) in 29 patients (15 D; 14 S). Propensity analysis and inverse probability of treatment weights (IPTW) to adjust for baseline differences were performed. Patients were followed for 2-7 years. Results - Avascular necrosis (AVN) occurred in 3 patients out of 15, after D, causing conversion to total hip replacement (THR) in 2 cases. In S, 1 hip developed chondrolysis, requiring THR 3 years after surgery. 3 symptomatic femoroacetabular impingements (FAI) occurred after S, requiring corrective osteotomy in 1 hip, and osteochondroplasty in another case. The risk of early re-operation was similar between the groups. The slippage was corrected more accurately and reliably by D. The Nonarthritic Hip Score was similar between groups, after adjusting for preoperative and postoperative variables. Interpretation - Although D was superior to S in restoring the proximal femoral anatomy, without increasing the risk of early re-operation, some concern remains regarding the potential risk of AVN in group D.


Characterization of a cis-acting regulatory element in the protein-coding region of human dihydrofolate reductase mRNA.
Previous studies have shown that human DHFR (dihydrofolate reductase), in addition to its critical role in DNA biosynthesis, functions as an RNA-binding protein. The interaction between DHFR and its own mRNA results in translational repression. In this study, we characterized the cis-acting elements on human DHFR mRNA that are required for the DHFR mRNA-DHFR protein interaction. Using a series of gel-shift and nitrocellulose filter-binding assays, a 164 nt RNA sequence, corresponding to nt 401-564, was identified within the coding region that binds to DHFR protein with an affinity similar to that of full-length DHFR mRNA. To document in vivo biological activity, various DHFR sequences contained within the coding region were cloned on to the 5' end of a luciferase reporter plasmid, and transient transfection experiments were performed using human colon cancer RKO cells. In cells transfected with p644/DHFR:401-564, luciferase activity was decreased by 50% when compared with cells transfected with the p644 plasmid alone. Luciferase mRNA levels were identical under each of these conditions, as determined by Northern-blot analysis. In cells transfected with p644/DHFR:401-564, luciferase activity was restored to almost 100% of control when cells were treated with the antifolate analogue methotrexate or with a short-interfering RNA targeting DHFR mRNA. These findings provide evidence that the DHFR 401-564 sequence is a DHFR-response element. In vitro and in vivo studies further localized this cis-element to an 82 nt sequence corresponding to nt 401-482. This work provides new insights into critical elements that mediate RNA-protein interactions.


Therapeutic targeting of the inflammasome in myeloid malignancies.
Even though genetic perturbations and mutations are important for the development of myeloid malignancies, the effects of an inflammatory microenvironment are a critical modulator of carcinogenesis. Activation of the innate immune system through various ligands and signaling pathways is an important driver of myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). The DAMPs, or alarmins, which activate the inflammasome pathway via the TLR4/NLR signaling cascade causes the lytic cell death of hematopoietic stem and progenitor cells (HSPCs), ineffective hematopoiesis, and β-catenin-induced proliferation of cancer cells, leading to the development of MDS/AML phenotype. It is also associated with other myeloid malignancies and involved in the pathogenesis of associated cytopenias. Ongoing research suggests the interplay of inflammasome mediators with immune modulators and transcription factors to have a significant role in the development of myeloid diseases, and possibly therapy resistance. This review discusses the role and importance of inflammasomes and immune pathways in myeloid malignancies, particularly MDS/AML, to better understand the disease pathophysiology and decipher the scope of therapeutic interventions.


Calcium- and otoferlin-dependent exocytosis by immature outer hair cells.
Immature cochlear outer hair cells (OHCs) make transient synaptic contacts (ribbon synapses) with type I afferent nerve fibers, but direct evidence of synaptic vesicle exocytosis is still missing. We thus investigated calcium-dependent exocytosis in murine OHCs at postnatal day 2 (P2)-P3, a developmental stage when calcium current maximum amplitude was the highest. By using time-resolved patch-clamp capacitance measurements, we show that voltage step activation of L-type calcium channels triggers fast membrane capacitance increase. Capacitance increase displayed two kinetic components, which are likely to reflect two functionally distinct pools of synaptic vesicles, a readily releasable pool (RRP; tau = 79 ms) and a slowly releasable pool (tau = 870 ms). The RRP size and maximal release rate were estimated at approximately 1200 vesicles and approximately 15,000 vesicles/s, respectively. In addition, we found a linear relationship between capacitance increase and calcium influx, like in mature inner hair cells (IHCs). These results give strong support to the existence of efficient calcium-dependent neurotransmitter release in immature OHCs. Moreover, we show that immature OHCs, just like immature IHCs, are able to produce regenerative calcium-dependent action potentials that could trigger synaptic exocytosis in vivo. Finally, the evoked membrane capacitance increases were abolished in P2-P3 OHCs from mutant Otof-/- mice defective for otoferlin, despite normal calcium currents. We conclude that otoferlin, the putative major calcium sensor at IHC ribbon synapses, is essential to synaptic exocytosis in immature OHCs too.


The asymmetry of chloride transport at 38 degrees C in human red blood cell membranes.
Band 3-mediated Cl- exchange in human red blood cells and resealed ghosts was measured at 38 degrees C by the continuous flow tube method. When external Cl- concentration, C(o), is varied with constant internal Cl- concentration, C(i), the flux fits a simple Michaelis-Menten saturation curve (MM fit), with K1/2o = 3.8 +/- 0.4 mM. When the Cl- concentration is varied simultaneously at both sides of the membrane in resealed ghosts (C(i) = C(o) = C(i = o)), the flux rises toward a flat maximum between 200 and 450 mM Cl-, and then decreases at very high C(i = o). An MM fit to the data with C(i = o) < 500 mM gives K1/2s of 106 +/- 13 mM; fits including modifier site inhibition (MS fit) give an over threefold higher K1/2s. Despite this uncertainty, the intrinsic asymmetry of unloaded transport sites, A (defined as E(o)/E(i) with C(i) = C(o), where E(i) is the fraction of unloaded inward-facing sites and E(o) is the fraction of unloaded outward-facing sites), calculated from K1/2s and K1/2o, ranges only from 0.046 to 0.107. A new method, which uses the initial slope of a plot of Cl- flux versus C(i = o), gives A values of 0.023 to 0.038. Flufenamic acid (FA) inhibits Cl- exchange by binding to an external site different from the transport site. At 38 degrees C, FA binds 24-36 times more tightly to E(o) than to E(i). Estimates of A from FA inhibitory potency range from 0.01 to 0.05. All methods, including bicarbonate data from the preceding paper, indicate that at 38 degrees C, like 0 degree C, far more band 3 molecules are in the E(i) than in the E(o) form. The agreement of various methods supports the ping-pong model for anion exchange, and demonstrates that the intrinsic asymmetry is very slightly, if at all, affected by temperature.


Aurora Borealis (Bora), Which Promotes Plk1 Activation by Aurora A, Has an Oncogenic Role in Ovarian Cancer.
Identifying novel actionable factors that critically contribute to tumorigenesis is essential in ovarian cancer, an aggressive and disseminative tumor, with limited therapeutic options available. Here we show that Aurora Borealis (BORA), a mitotic protein that plays a key role in activating the master mitotic kinase polo-like kinase 1 (PLK1), has an oncogenic role in ovarian cancer. Gain and loss of function assays on mouse models and ex vivo patient-derived ascites cultures revealed an oncogenic role of BORA in tumor development and a transcriptome-analysis in clinically representative models depicted BORA's role in survival, dissemination and inflammatory cancer related-pathways. Importantly, combinatory treatments of FDA-approved inhibitors against oncogenic downstream effectors of BORA displayed synergistic effect in ovarian cancer models, offering promising therapeutic value. Altogether, our findings uncovered for the first time a critical role of BORA in the viability of human cancer cells providing potential novel therapeutic opportunities for ovarian cancer management.


Spontaneous calcification process in primary renal cells from a medullary sponge kidney patient harbouring a GDNF mutation.
Medullary nephrocalcinosis is a hallmark of medullary sponge kidney (MSK). We had the opportunity to study a spontaneous calcification process in vitro by utilizing the renal cells of a patient with MSK who was heterozygous for the c.-27 + 18G>A variant in the GDNF gene encoding glial cell-derived neurotrophic factor. The cells were obtained by collagenase digestion of papillary tissues from the MSK patient and from two patients who had no MSK or nephrocalcinosis. These cells were typed by immunocytochemistry, and the presence of mineral deposits was studied using von Kossa staining, scanning electron microscopy analysis and an ALP assay. Osteoblastic lineage markers were studied using immunocytochemistry and RT-PCR. Staminality markers were also analysed using flow cytometry, magnetic cell separation technology, immunocytochemistry and RT-PCR. Starting from p2, MSK and control cells formed nodules with a behaviour similar to that of calcifying pericytes; however, Ca₂PO₄ was only found in the MSK cultures. The MSK cells had morphologies and immunophenotypes resembling those of pericytes or stromal stem cells and were positive for vimentin, ZO1, αSMA and CD146. In addition, the MSK cells expressed osteocalcin and osteonectin, indicating an osteoblast-like phenotype. In contrast to the control cells, GDNF was down-regulated in the MSK cells. Stable GDNF knockdown was established in the HK2 cell line and was found to promote Ca₂PO₄ deposition when the cells were incubated with calcifying medium by regulating the osteonectin/osteopontin ratio in favour of osteonectin. Our data indicate that the human papilla may be a perivascular niche in which pericyte/stromal-like cells can undergo osteogenic differentiation under particular conditions and suggest that GDNF down-regulation may have influenced the observed phenomenon.


Cardiovascular risks of COX inhibition: current perspectives.
NSAIDs are among the most commonly used pharmacotherapeutic agents worldwide. As the long-term use of these drugs is associated with serious gastrointestinal side effects, a new subgroup of COX-2 selective NSAIDs was developed. It was thought that the therapeutic strategy underlying the development of these newer compounds would enable them to provide the same analgesic and anti-inflammatory benefits as those of their traditional counterparts but perhaps offer a much safer gastrointestinal profile. Much scientific data has accumulated over the last few years, however, raising concerns regarding the increased cardiovascular complications associated with the use of COX-2 selective NSAIDs, and perhaps of the traditional NSAIDs as well. To review current and emerging evidence related to the cardiovascular effects of COX inhibitors and examine the clinical implications. We studied data from basic clinical research, non-randomized analyses, and randomized trials of COX inhibitors that investigated their cardiovascular effects. Both COX-2 selective and traditional NSAIDs are associated with a moderately increased risk of cardiovascular events.


Primary omental torsion: case report.
Primary omental torsion is a rare condition. It is confused with acute appendicitis. Presence of a normal appendix at surgery should raise the suspicion of omental torsion.


[Choice of anesthesia in patients with suppurative surgical infections and diabetes mellitus].
500 anesthesias were performed in patients with surgical diseases in the presence of diabetes millitus (90% of patients with II type diabetes). The methods of intravenous anesthesia are proposed for short-term procedures with preservation of spontaneous ventilation and stable hemodynamics. The method of epidural anesthesia with introduction of major dose of anesthetic in two stages was developed for the operation on the lower extermities, it provides for stable hemodynamics and adequate anesthesia. Factors, influencing the choice of anesthesia were determined. Basic principles of intensive care in patients with surgical infection in diabetes mellitus were established.


Injury surveillance in Trinidad: an accident and emergency based injury surveillance system at the San Fernando General Hospital.
Injuries are a significant cause of morbidity and mortality worldwide. Injuries disproportionately affect people living in low and middle income countries, including the Caribbean; however little is known about the epidemiology of injuries in these areas. An accident and emergency (A&E) department injury surveillance system was established at the San Fernando General Hospital, Trinidad and Tobago, to address this important data gap. A detailed overview of the objectives, data collection methods, and inherent strengths and limitations of this surveillance system are presented, along with results of an analysis of data collected during the first three years of operations (from 2002 to 2004). Trained hospital staff collect a variety of injury/poisoning, demographic and clinical data on nearly 20,000 patients presenting each year with injury to the A&E Department. The total number of injuries in men was almost twice that in women. The majority of injuries were seen in those 25-44-years of age. Falls, other blunt force, stab/cut, traffic injury and poisoning represented the leading causes of injury. Nearly half of all the injuries occurred in the home, with the street/highway and work environments also accounting for an appreciable number of injuries. The majority of injuries were reported as unintentional. Injuries represent an important population health and health services issue in South Trinidad. Data from the A&E Department injury surveillance system represent an important resource to inform evidence-based health policy decisions on injury prevention and public health resource allocation.


Interleukin-7 restores immunity in athymic T-cell-depleted hosts.
Thymic-deficient hosts rely primarily on antigen-driven expansion to restore the peripheral T-cell compartment following T-cell depletion (TCD). The degree to which this thymic-independent pathway can restore immune competence remains poorly understood but has important implications for a number of clinical conditions including stem cell transplantation and human immunodeficiency virus (HIV) infection. A model of HY-mediated skin graft rejection by athymic, TCD mice was used to show that restoration of naive and recall responses via peripheral expansion requires transfer of only 25 x 10(6) lymph node (LN) cells representing approximately 10% of the T-cell repertoire. Constitutive expression of bcl-2 in the expanding inocula restored recall responses to HY at a substantially lower LN cell dose (1 x 10(6)), which is normally insufficient to induce HY-mediated graft rejection in athymic hosts. Interestingly, bcl-2 had no effect on primary responses. Interleukin-7 (IL-7) potently enhanced thymic-independent peripheral expansion and led to HY graft rejection using an LN cell dose of 1 x 10(6) in both primary and recall models. The restoration of immune competence by IL-7 appeared to be mediated through a combination of programmed cell death inhibition, improved costimulation, and modulation of antigen-presenting cell (APC) function. These results show that immune competence for even stringent antigens such as HY can be restored in the absence of thymic function and identify IL-7 as a potent modulator of thymic-independent T-cell regeneration.


Beyond DNA repair and chromosome instability-Fanconi anaemia as a cellular senescence-associated syndrome.
Fanconi anaemia (FA) is the most frequent inherited bone marrow failure syndrome, due to mutations in genes encoding proteins involved in replication fork protection, DNA interstrand crosslink repair and replication rescue through inducing double-strand break repair and homologous recombination. Clinically, FA is characterised by aplastic anaemia, congenital defects and cancer predisposition. In in vitro studies, FA cells presented hallmarks defining senescent cells, including p53-p21 axis activation, altered telomere length, mitochondrial dysfunction, chromatin alterations, and a pro-inflammatory status. Senescence is a programme leading to proliferation arrest that is involved in different physiological contexts, such as embryogenesis, tissue remodelling and repair and guarantees tumour suppression activity. However, senescence can become a driving force for developmental abnormalities, aging and cancer. Herein, we summarise the current knowledge in the field to highlight the mutual relationships between FA and senescence that lead us to consider FA not only as a DNA repair and chromosome fragility syndrome but also as a "senescence syndrome".


Is there an alternative to a National Health Board for Technology Assessment?
The National Health Board, a lynchpin of the managed competition philosophy of health reform, would perform a necessary function in regards to technology assessment for clinical decision making. However, would such an agency have what it takes to provide adequate turnaround, and nonbiased decision-making ability to play this crucial part of the reform process?


Impurity-driven nanocrystallization of Zr-based bulk amorphous alloys.
The effect of oxygen content and Ti addition on the glass forming ability (GFA) and crystallization kinetic of Zr-based bulk glass forming alloys have been studied by means of thermal analysis and X-ray diffraction techniques. Presence of oxygen triggers the formation of a nanocrystalline metastable f.c.c. Zr2 Ni-type phase which can act as heterogenous nucleation sites for the formation of dendrites during solidification. An increase in oxygen content changes the crystallization behaviour from a single- to a double-step process and triggers the crystallization of stable Zr2(Cu,Al) besides metastable Zr2 Ni-type phase. Oxygen-triggered nucleation of a nanocrystalline metastable Zr2 Ni-type phase is found to be the initial step of crystallization. The important parameters of GFA such as glass transition temperatures, Tg, the crystallization temperatures, Tx, and crystallization enthalpies, deltaH, were determined by using DSC. It was observed that the presence of oxygen, even in a very small amount, and Ti addition cause a drastic reduction of the supercooled liquid region, deltaTx, accompanied by a change of the crystallization kinetic. This leads to the decrease in stability of the supercooled liquid, consequently results in a deterioration of the glass forming ability of the alloy.


The Spectrum of Molecular Pathways in Gliomas-An Up-to-Date Review.
During the last 20 years, molecular alterations have gained increasing significance in the diagnosis and biological assessment of tumors. Gliomas represent the largest group of tumors of the central nervous system, and the main aim of this review is to present the current knowledge on molecular pathways and their alterations in gliomas. A wide range of new insights has been gained, including evidence for the involvement of the WNT pathway or the hippo pathway in the pathobiology of gliomas, indicating a broad involvement of different pathways formerly not considered to play a central role in gliomas. Even new aspects of angiogenic, apoptotic, and metabolic pathways are presented, as well as the rapidly growing field of epigenetic processes, including non-coding RNAs. The two major conclusions drawn from the present review are the distinct interconnectivity of the whole spectrum of molecular pathways and the prominent role of non-coding RNAs, especially circular RNAs, in the regulation of specific targets. All these new insights are discussed, even considering the topic of the resistance to therapy of gliomas, along with aspects that are still incompletely understood, like the role of hydroxymethylation, or even ferroptosis, in the pathobiology of gliomas.


Tumor immunoediting by NKp46.
NK cells interact with a wide variety of hazardous cells including pathogen-infected and tumor cells. NKp46 is a specific NK killer receptor that recognizes various influenza hemagglutinins and unknown tumor ligands. It was recently shown that NKp46 plays a significant role in the in vivo eradication of tumor cells; however, the role played by NKp46 in vivo with regard to tumor development is still unclear. In this study, we used the 3-methylcholanthrene (MCA)-induced fibrosarcoma model in NKp46-deficient mice to test the NKp46 recognition of carcinogen-induced tumors. We show that although the rate of MCA-induced tumor formation was similar in the presence and in the absence of NKp46, the expression of its unknown ligands was NKp46 dependent. The unknown NKp46 ligands were nearly absent in tumors that originated in wild-type mice, whereas they were detected in tumors that originated in the NKp46-deficient mice. We demonstrate that the interactions between NKp46 and its MCA tumor-derived ligands lead to the secretion of IFN-gamma but not to the elimination of the MCA-derived tumor cells. In addition, we show that the in vivo growth of MCA-derived tumor cells expressing high levels of the NKp46 ligands is NKp46 and IFN-gamma dependent. Thus, we present in this study a novel NKp46-mediated mechanism of tumor editing.


Symptoms of estrogen deficiency in nursing personnel in Maharaj Nakorn Chiang Mai Hospital.
To determine the self-reported prevalence and severity of climacteric symptoms of estrogen deficiency in nursing personnel working in Maharaj Nakorn Chiang Mai Hospital during the year 2002. Cross-sectional descriptive study. Department of Obstetrics and Gynecology, Faculty of Medicine, Chiang Mai University. Six-hundred and eight nursing personnel, including professional nurses and practical nurses, aged 40-60 years, working in Maharaj Nakorn Chiang Mai Hospital. The same sets of questionnaires were provided and distributed to each subject. Subjects had self assessment under individual decision with written consent and data was prospectively collected. The data were analyzed using software SPSS version 10 and presented in frequencies and percentage. The prevalence and severity of each climacteric symptoms. Five-hundred and seventy five of 608 subjects (94.6%) responded to the questionnaires, 62.6% of them were in normal reproductive period, while the remainder (37.4%) were in the menopausal period. The latter group was divided into 4 subgroups of pre/peri-menopause, post menopause, surgical menopause and premature ovarian failure. The prevalence of climacteric symptoms was systematically classified as follows, vasomotor instability 40.7% (severe cases, 5.4%), psychosomatic symptoms 50.9% (severe cases, 3.9%), lower urinary tract symptoms 29.1% (severe cases, 2.4%), lower genital tract symptoms 34.0% (severe cases, 2.7%), and other symptoms 50.7% (severe cases, 4.3%). The first five most common symptoms were as follows, forgetful 84.1% (severe cases, 7.9%), myalgia 74.3% (severe cases, 10.6%), anxious 71.0% (severe cases, 5.3%), tired 70.0% (severe cases, 4.1%), headache 68.3% (severe cases, 8.3%). The least common symptom was needle pain, the prevalence was 19.7% (severe cases, 1.3%). The overall prevalence of women using hormone replacement therapy was 13.1%, only 8.9% were current users. Focusing on each symptom of climacteric symptoms, the authors found high prevalence in psychosomatic symptoms and other symptoms such as forgetful, myalgia, anxious, tired and headache. While the classic symptom (vasomotor instability) was found to be low in the present study which was different from the previous reports. This may be due to the racial or cultural and educational factors among different populations.


The negative inotropic action of catecholamines: role of beta3-adrenoceptors.
There is now evidence for the involvement of four beta-adrenoceptor populations in the regulation of cardiac function by catecholamines. Beta1- and beta2-adrenoceptor stimulation classically produces an increase in contractility. A fourth beta-adrenoceptor, as yet uncloned and designated provisionally as a beta4-adrenoceptor, also mediates a positive inotropic effect. Beta3-adrenoceptors, which had been cloned at the end of the eighties, has been extensively studied as a potential target for antiobesity and antidiabetic drugs. Its characterization in the heart has opened new fields of investigations for the understanding of the cardiac adrenergic regulation. This review describes the cardiac electrical and mechanical effects induced by Beta3-adrenoceptor stimulation in different species (including human), as well as the signaling pathway. It also analyzes the role of these receptors in the abnormal responsiveness of catecholamines in heart failure.


Tuning Escherichia coli for membrane protein overexpression.
A simple generic method for optimizing membrane protein overexpression in Escherichia coli is still lacking. We have studied the physiological response of the widely used "Walker strains" C41(DE3) and C43(DE3), which are derived from BL21(DE3), to membrane protein overexpression. For unknown reasons, overexpression of many membrane proteins in these strains is hardly toxic, often resulting in high overexpression yields. By using a combination of physiological, proteomic, and genetic techniques we have shown that mutations in the lacUV5 promoter governing expression of T7 RNA polymerase are key to the improved membrane protein overexpression characteristics of the Walker strains. Based on this observation, we have engineered a derivative strain of E. coli BL21(DE3), termed Lemo21(DE3), in which the activity of the T7 RNA polymerase can be precisely controlled by its natural inhibitor T7 lysozyme (T7Lys). Lemo21(DE3) is tunable for membrane protein overexpression and conveniently allows optimizing overexpression of any given membrane protein by using only a single strain rather than a multitude of different strains. The generality and simplicity of our approach make it ideal for high-throughput applications.


Estimation of neural noise. Functional anatomy of the human thalamus.
In the course of stereotactic thalamotomy, the neural noise level of subcortical structures was estimated quantitatively with the aid of two semimicro-electrodes. The neural noise showed several characteristic features in terms of its amplitude and discharge pattern so that it was correlated with the possible anatomical substrate, thus providing the functional anatomy of the subcortical structures. The study on saggital plane revealed a systematic difference of the noise level between VL and Vim-Vc that could be explained by the different cell size in respective nucleus. Several exceptional cases were also presented, emphasizing the neural noise study in stereotactic surgery.


Work-related factors associated with occupational exposure to static magnetic stray fields from MRI scanners.
This study aims to identify work-related and personal factors associated with workers' exposure to static magnetic fields (SMF) and motion-induced time-varying magnetic fields (TVMF) from MRI scanners. Measurements of personal exposure to SMF and TVMF were performed among MRI staff during 439 work shifts at 14 different workplaces using portable magnetic field dosimeters. These data were coupled with contextual workplace and worker information. After data cleanup, 324 remaining observations were used to develop linear mixed effects models for various measures of peak and time-weighted average (TWA) exposure. Exposure levels near whole-body closed-bore scanners increased by 30% to 76% for each additional tesla of scanner strength, depending on exposure metric. Small-bore animal scanners, on the other hand, showed a reversed association with scanner strength. Measures of peak and TWA exposure were differently associated with specific tasks and scan procedures. In addition, body height of the worker was negatively associated with measured exposure levels. The study revealed workplace characteristics, scan activities, and personal characteristics associated with SMF and TVMF exposure levels of MRI staff and was able to quantify the unique contribution of each of these factors while adjusting for the presence of others.


Presence and influence of cholinergic nerves in the human thyroid.
There is evidence that the sympathetic nervous system exerts a control on human thyroid function via an adrenergic innervation of follicle cells. The present study demonstrates that cholinergic nerve fibers also reach follicle cells in the normal human thyroid. In addition, cholinergic agents were found to enhance cGMP accumulation in human thyroid tissue. This effect was blocked by atropine, a muscarinic receptor antagonist, but not by d-tubocurarine, a nicotinic receptor antagonist. These results provide morphological and biochemical arguments supporting a role of the parasympathetic nervous system in the regulation of thyroid function in man.


[Dysphagia associated with aorto-cardiac compression on the distal esophagus: a rare event but not exceptional in the elderly].
Among the pathologies responsible for dysphagia in the elderly, the aortic compression on the distal oesophagus awakens a certain clinical interest. Cipho-lordosis, which is often present in aged females, worsens the effects of this compression causing other kinkings on the oesophagus already hypokinetic because of the age (presbyoesophagus). This illness often shows scare symptoms, but sometimes requires a pneumatic dilation to allow normal food assumption of the patient. The authors report on their experience of two cases, clearly documented from the diagnostic point of view; the first one was given medical treatment, the second one was treated with a pneumatic dilatation of the cardias.


Disparities in brachytherapy utilization in cervical cancer in the United States: A comprehensive literature review.
Brachytherapy (BT) is an integral component of treatment for patients with locally advanced cervical cancer, significantly improving local control and overall survival. There is an overall trend of decreased utilization of BT in United States (US) in the last few decades with around 50% of patients being treated without BT. The cause of decreased utilization is multifactorial including physician comfort, facility volume, low reimbursements rates and costs of starting and maintaining a brachytherapy program. This decrease coincides with an increase in the use of newer advanced techniques like intensity modulated radiation therapy (IMRT) and stereotactic body radiation therapy (SBRT) boost resulting in inferior oncological outcomes and increased toxicity. Moreover, racial and socioeconomic disparities in BT utilization have been widely reported in the US. Various factors including age, race, socio-economic status, location, facility type, facility volume and insurance status result in limited access to brachytherapy, which jeopardizes oncologic outcomes. This comprehensive review discusses the BT utilization in the US, examines the impact of race and socioeconomic factors on BT utilization, and highlights its impact on outcomes.


Defining the human C2H2 zinc finger degrome targeted by thalidomide analogs through CRBN.
The small molecules thalidomide, lenalidomide, and pomalidomide induce the ubiquitination and proteasomal degradation of the transcription factors Ikaros (IKZF1) and Aiolos (IKZF3) by recruiting a Cys2-His2 (C2H2) zinc finger domain to Cereblon (CRBN), the substrate receptor of the CRL4CRBN E3 ubiquitin ligase. We screened the human C2H2 zinc finger proteome for degradation in the presence of thalidomide analogs, identifying 11 zinc finger degrons. Structural and functional characterization of the C2H2 zinc finger degrons demonstrates how diverse zinc finger domains bind the permissive drug-CRBN interface. Computational zinc finger docking and biochemical analysis predict that more than 150 zinc fingers bind the drug-CRBN complex in vitro, and we show that selective zinc finger degradation can be achieved through compound modifications. Our results provide a rationale for therapeutically targeting transcription factors that were previously considered undruggable.


[Pediatric pharmacology: is treatment of children preceded by an adequate investigation?].
The articles published in 1985 in 9 journals (3 of Internal Medicine, 3 of Pediatrics and 3 of Clinical Pharmacology) are revised with the purpose of studying: a) Number of clinical trials published. b) Number of clinical trials published accomplished in children. Among the 533 trials published, 102 were done in children. The type of trial done is studied based on the methodology used, if healthy volunteers are included, if pharmacokinetics studies were done, the ages of the subjects, the number of subjects included, the sex and the way of obtaining the informed consent.


Paclitaxel and G-CSF in previously untreated patients with extensive stage small-cell lung cancer: a phase II study of the North Central Cancer Treatment Group.
Paclitaxel is an antimicrotubule agent that interferes with cell division. It has demonstrated promising single-agent activity against non-small-cell lung cancer. The purpose of this study was to evaluate the therapeutic effectiveness of paclitaxel in previously untreated patients with extensive stage small-cell lung cancer (SCLC). The study was designed as a two-stage phase II trial. All patients who entered received paclitaxel by intravenous infusion at a dose of 250 mg/m2 during 24 hours. Granulocyte colony stimulating factor was also provided to ameliorate neutropenia. Cycles were repeated at 21-day intervals. Patients who achieved a complete response received a maximum of 10 cycles of treatment, whereas those who achieved a partial response/regression continued treatment until progression or undue toxicity developed. Patients who progressed or maintained stable disease for six cycles were crossed over to cisplatin and etoposide. Forty-three patients entered the study and all were evaluable for analysis. Responses were observed in 23 (53%) of the patients. There was no significant difference in the response rates in patients with measurable or evaluable disease (13/23 versus 10/20, p = 0.76). At the time of analysis, 39 patients had progressed with a median time to progression of 95 days, and 39 patients had died with a median survival of 278 days. The 1-year achieved survival rate was 24%. Significant neutropenia (absolute neutrophil count <1,000/microl) occurred in 24 (56%) of the patients, but only 2 patients experienced severe infection (grade > or = 3), and there were no septic deaths. The results indicate that paclitaxel is active against SCLC. Myelosuppression was the main side effect in this patient population. Response duration was short (median = 3.4 months), which suggests that paclitaxel is not sufficient as a single agent. Further studies of paclitaxel in combination with other agents against SCLC are currently in progress within the North Central Cancer Treatment Group and other cancer treatment groups. Key Words: Paclitaxel-G-CSF-Small-cell lung cancer-North Central Cancer Treatment Group.


HPV testing in combination with liquid-based cytology in primary cervical screening (ARTISTIC): a randomised controlled trial.
Testing for human papillomavirus (HPV) DNA is reportedly more sensitive than cytology for the detection of high-grade cervical intraepithelial neoplasia (CIN). The effectiveness of HPV testing in primary cervical screening was assessed in the ARTISTIC trial, which was done over two screening rounds approximately 3 years apart (2001-03 and 2004-07) by comparing liquid-based cytology (LBC) combined with HPV testing against LBC alone. Women aged 20-64 years who were undergoing routine screening as part of the English National Health Service Cervical Screening Programme in Greater Manchester were randomly assigned (between July, 2001, and September, 2003) in a ratio of 3:1 to either combined LBC and HPV testing in which the results were revealed and acted on, or to combined LBC and HPV testing where the HPV result was concealed from the patient and investigator. The primary outcome was the detection rate of cervical intraepithelial neoplasia grade 3 or worse (CIN3+) in the second screening round, analysed by intention to treat. This trial is registered with the International Standard Randomised Controlled Trial Number ISRCTN25417821. There were 24 510 eligible women at entry (18 386 in the revealed group, 6124 in the concealed group). In the first round of screening 233 women (1.27%) in the revealed group had CIN3+, compared with 80 (1.31%) women in the concealed group (odds ratio [OR] 0.97, 95% CI 0.75-1.25; p>0.2). There was an unexpectedly large drop in the proportion of women with CIN3+ between the first and second rounds of screening in both groups, at 0.25% (29 of 11 676) in the revealed group and 0.47% (18 of 3866 women) in the concealed group (OR 0.53, 95% CI 0.30-0.96; p=0.042). For both rounds combined, the proportion of women with CIN3+ were 1.51% (revealed) and 1.77% (concealed) (OR 0.85, 95% CI 0.67-1.08; p>0.2). LBC combined with HPV testing resulted in a significantly lower detection rate of CIN3+ in the second round of screening compared with LBC screening alone, but the effect was small. Over the two screening rounds combined, co-testing did not detect a higher rate of CIN3+ or CIN2+ than LBC alone. Potential changes in screening methodology should be assessed over at least two screening rounds. National Institute of Health Research Health Technology Assessment Programme.


TLR7 agonist administration to SIV-infected macaques receiving early initiated cART does not induce plasma viremia.
Reduction/elimination of HIV-1 reservoirs that persist despite combination antiretroviral therapy (cART) will likely require induction of viral expression by residual infected cells and enhanced clearance of these cells. TLR7 agonists have potential to mediate these activities. We evaluated immunologic and virologic effects of repeated doses of the TLR7 agonist GS-9620 in SIV-infected rhesus macaques receiving cART, which was initiated at 13 days after infection and was continued for 75 weeks prior to GS-9620 administration. During cART, GS-9620 induced transient upregulation of IFN-stimulated genes in blood and tissues, increases in plasma cytokines, and changes in immune cell population activation and phenotypes but did not result in measurable increases in plasma viremia or viral RNA-to-viral DNA ratio in PBMCs or tissues nor decreases in viral DNA in PBMC or tissues. SIV-specific CD8+ T cell responses, negligible prior to GS-9620 treatment, were not measurably boosted by treatment; a second course of GS-9620 administration overlapping with later cART discontinuation was associated with increased CD8+ T cell responses during viral recrudescence. These results confirm and extend evidence for GS-9620-mediated enhancement of antiviral immune responses in SIV-infected macaques but suggest that GS-9620-mediated viral induction may depend critically on the timing of initiation and duration of cART and resulting characteristics of viral reservoirs.


Association of MHC antigens with susceptibility to and severity of rheumatoid arthritis in multicase families.
A study of HLA association with rheumatoid arthritis (RA) in multicase families has been performed in north east England. Two hundred and nineteen individuals from 13 families were assessed for the presence of RA, and all were HLA typed. Thirty-nine were found to have classical or definite RA by American Rheumatism Association (ARA) criteria. Thirty-five (90%) of these possess HLA-DR4, confirming the previously reported association of RA with DR4. A further 19 individuals were found to have probable RA or gave a convincing history of previous inflammatory polyarthritis. Thirteen (68%) of these possess HLA-DR4, and this is not significantly different from non-affected family members of whom 63% possess DR4. These results suggest that HLA-DR4 is associated only with the more severe forms of RA. Homozygosity for HLA-DR4 was not associated with either earlier onset or more severe disease when compared with heterozygous DR4. Possession of the haplotype most commonly inherited with the RA in individual families was not associated with earlier onset but may be associated with more severe disease. The severity of RA appears to be influenced by the major histocompatibility complex (MHC) in these families.


Craniofacial growth in children treated for malignant diseases.
With the improving cure rate in childhood malignancies, increasing interest has been focused on the long-term survivors of childhood cancer and the quality of their life. The severity of long-term disturbances in dental and craniofacial development is dependent on the age of the child at diagnosis, if chemotherapy is combined with radiation or not. With regard to craniofacial development combination chemotherapy has no effects compared with healthy controls, whereas children treated cranial irradiation before 5 years of age exhibit a reduced growth of the mandible. Conditioning before bone marrow transplantation with total body irradiation results in a significantly reduced growth of the craniofacial skeleton. The mandible was four times more radiosensitive compared with the maxilla. With attention to the dental and craniofacial development, occlusion and craniomandibular function, children in risk groups should be followed, and given prophylactic treatment and intervention at appropriate times to reduce the consequences of the disease itself and the therapy given.


Hepatocellular carcinoma: combined hepatic, arterial, and portal venous embolization.
Transcatheter arterial embolization (TAE) is an effective means of treating primary hepatocellular carcinoma (HCC). However, in many cases of HCC the tumor recurs after treatment. In an attempt to obtain complete tumor necrosis, the authors studied the clinical and histologic effect of simultaneous embolization of both the hepatic artery and portal vein in ten patients with HCC. In those cases in which combined embolization caused infarction, tumor cells in the main tumor, tumor cells that had invaded the tumor capsule, and small intrahepatic metastases had become totally necrotic following treatment. No viable tumor cells were detected in four patients who subsequently underwent operations; nor were viable tumor cells present in one other patient who later died as a result of a perforated duodenal ulcer. Five patients who did not subsequently undergo operations were still free of the disease 2-17 months after combined arterial and portal embolization. The impact of combined embolization on liver function was nearly the same as that produced when TAE was performed alone. Combined embolization may be a viable alternative to hepatectomy for the treatment of HCC.


Long-term combined supplementations with alpha-tocopherol and vitamin C have no detectable anti-inflammatory effects in healthy men.
Inflammatory and oxidative stresses play a pivotal role in atherogenesis. Vitamin E and vitamin C are the two most important dietary antioxidants; moreover, vitamin E has anti-inflammatory effects. Combined supplementations with vitamin E and vitamin C twice daily for 3 y reduced lipid peroxidation and retarded the progression of common carotid atherosclerosis in healthy men in the Antioxidant Supplementation in Atherosclerosis Prevention (ASAP) study. To further elucidate the underlying mechanisms that retarded the progression of atherosclerosis in the ASAP study, we investigated the effect of a combined intake of vitamin E and vitamin C on inflammatory markers in vivo. Circulating levels of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and C reactive protein (CRP) were measured in 45- to 69-y-old men from the ASAP study with cholesterol >5.0 mmol/L before and after treatment with either placebo (n = 52) or a combined supplementation with 91 mg (136 IU) alpha-tocopherol and 250 mg of slow-release vitamin C twice a day (n = 55) for 3 y. Antioxidant treatment for 36 mo had no effect on circulating levels of TNF-alpha, IL-6 or CRP. In conclusion, long-term combined supplementations with alpha-tocopherol and vitamin C in reasonable doses have no detectable systemic anti-inflammatory effects in a healthy population of men with slight hypercholesterolemia and no overt signs of inflammation.


The Ethical Digital Surgeon.
This viewpoint explores the ethical and regulatory consequences of the digital transformation of the operating room. Surgical robotics is undergoing significant change and future advances will center around the capture and use of data. The consequences of creating this surgical data pipeline must be understood and digital surgical systems must prioritize the safeguarding of patient data. Moreover, data protection laws and frameworks must adapt to the changing nature of surgical data. Finally, digital surgeons must understand changing data legislation and best practice on data governance to act as guardians not only for their own but also for their patients' data.


Chlamydia pneumoniae inhibits activated human T lymphocyte proliferation by the induction of apoptotic and pyroptotic pathways.
Chlamydia pneumoniae is an omnipresent obligate intracellular bacterial pathogen that infects numerous host species. C. pneumoniae infections of humans are a common cause of community acquired pneumonia but have also been linked to chronic diseases such as atherosclerosis, Alzheimer's disease, and asthma. Persistent infection and immune avoidance are believed to play important roles in the pathophysiology of C. pneumoniae disease. We found that C. pneumoniae organisms inhibited activated but not nonactivated human T cell proliferation. Inhibition of proliferation was pathogen specific, heat sensitive, and multiplicity of infection dependent and required chlamydial entry but not de novo protein synthesis. Activated CD4(+) and CD8(+) T cells were equally sensitive to C. pneumoniae antiproliferative effectors. The C. pneumoniae antiproliferative effect was linked to T cell death associated with caspase 1, 8, 9, and IL-1β production, indicating that both apoptotic and pyroptotic cellular death pathways were activated after pathogen-T cell interactions. Collectively, these findings are consistent with the conclusion that C. pneumoniae could induce a local T cell immunosuppression and inflammatory response revealing a possible host-pathogen scenario that would support both persistence and inflammation.


Metabolic effects of lathyrogenic agents on cartilage in vivo and in vitro.
The effects of lathyrogenic agents in vivo and in vitro are described, in respect to some biochemical indices of cartilage metabolism. Lathyrogenic agents in vivo inhibited the incorporation of radiosulfate into rat epiphyseal cartilage and the isolated chondroitin sulfate. No significant changes in hydroxyproline or hexosamine content of epiphyseal cartilage were found, but there was a marked increase in water content. The content of chondroitin sulfate, measured as uronic acid, was decreased. The importance of taking growth rate differences between control and experimental rats into account in assessing the effects of lathyrogenic agents in vivo is emphasized. In an in vitro system, utilizing fresh calf costal cartilage slices, the presence of low concentrations of lathyrogenic agents markedly affected various metabolic events. The incorporation into cartilage slices of sulfate-S(35), glucose-U-C(14), and glycine-1-C(14) was significantly depressed, as was the production of organic acids, including lactic acid. In general, these effects were more severe under anaerobic conditions. Glutamine restored the activities of the slices treated with lathyrogenic agents to control values obtained in the absence of either lathyrogen or glutamine.


Cargo adapters expand the transport range of intraflagellar transport.
The assembly and maintenance of most cilia and eukaryotic flagella depends on intraflagellar transport (IFT), the bidirectional movement of multi-megadalton IFT trains along the axonemal microtubules. These IFT trains function as carriers, moving ciliary proteins between the cell body and the organelle. Whereas tubulin, the principal protein of cilia, binds directly to IFT particle proteins, the transport of other ciliary proteins and complexes requires adapters that link them to the trains. Large axonemal substructures, such as radial spokes, outer dynein arms and inner dynein arms, assemble in the cell body before attaching to IFT trains, using the adapters ARMC2, ODA16 and IDA3, respectively. Ciliary import of several membrane proteins involves the putative adapter tubby-like protein 3 (TULP3), whereas membrane protein export involves the BBSome, an octameric complex that co-migrates with IFT particles. Thus, cells employ a variety of adapters, each of which is substoichiometric to the core IFT machinery, to expand the cargo range of the IFT trains. This Review summarizes the individual and shared features of the known cargo adapters and discusses their possible role in regulating the transport capacity of the IFT pathway.


Effectiveness of bonded and vacuum-formed retainers: A prospective randomized controlled clinical trial.
The objective of this prospective trial was to compare the clinical effectiveness of bonded retainers with vacuum-formed retainers, in terms of maintaining the results of orthodontic treatment in the lower arch up to 18 months post debond. This was a hospital-based, prospective randomized controlled clinical trial in which a total of 82 subjects were randomly allocated using a computer-generated number sequence to 1 of 2 groups, receiving either a vacuum-formed retainer (Essix Ace plastic (120 mm; DENTSPLY Raintree Essix, Sarasota, Fla) or a bonded retainer (0.0175 coaxial archwire (Ortho-Care, UK, Shipley, United Kingdom) bonded in place with Transbond LR (3M United Kingdom, Brachnell, United Kingdom) for the mandibular arch. Each number was placed in an opaque, concealed envelope and chosen randomly by the study subject; this determined the allocation group. Eligibility criteria included patients nearing debond after treatment with 0.022 × 0.028-in slot size preadjusted edgewise fixed orthodontic appliances whose pretreatment records and study models were available to confirm pretreatment labial segment crowding or spacing and who had clinically acceptable alignment at the end of treatment. The main outcome was to investigate the clinical effectiveness of the 2 types of retainers in terms of changes in incisor irregularity at 6 months of retention. The following measurements were recorded at each time point (6, 12, and 18 months) with a digital caliper: Little's irregularity index, intercanine width, intermolar width, arch length, and extraction site opening. Blinding was applicable only at debond because of the permanence of 1 intervention. The 2 groups were well matched with respect to age, sex, clinical characteristics, and treatment plans. There was a statistically significant difference between the groups for changes in Little's irregularity index at 6 months, with the vacuum-formed retainer group showing greater changes than the bonded retainer group (P = 0.008). There was no statistically significant difference between the groups for changes in Little's irregularity index at 12 and 18 months.There were also no statistically significant changes at any time for intercanine width, intermolar width, arch length, or extraction site opening. Some relapse is likely after fixed appliance therapy irrespective of retainer choice, and this is minimal in most patients at 6 months after debond. Bonded retainers have a better ability to hold the mandibular incisor alignment in the first 6 months after treatment than do vacuum-formed retainers. Not applicable. The protocol was not published before trial commencement. There is no funding or conflict of interest to be declared.


[ T- and B-lymphocyte populations in thyroid pathology].
Examination of cellular and humoral immunity in patients with thyroid pathology demonstrated that autoimmune forms were attended by T cell deficiency with an excess of B cells and circulating antithyroid autoantibodies. The blood content of T and B lymphocytes was within the normal range in patients with nontoxic goiter, but circulating autoantibodies to the thyroid gland were found in 40% of cases. Examination of the blood in patients with diffuse toxic goiter of moderate severity at the early periods after a course of chemotherapy showed no normalization of the cellular and humoral immunity indices.


Epidermal growth factor induces coupling of protein-tyrosine phosphatase 1D to GRB2 via the COOH-terminal SH3 domain of GRB2.
The Src homology 2 (SH2) and SH3 domain-containing adaptor protein GRB2 and the SH2 domain-containing protein-tyrosine phosphatase 1D (PTP1D, also called SHPTP2, PTP2C, SHPTP3, Syp, or SHP-2) function as positive mediators of growth factor-induced mitogenesis. Epidermal growth factor (EGF) is a potent mitogen for MCF-10A human mammary epithelial cells and EGF receptor-expressing mouse NR6 fibroblasts. Western blot analysis of anti-PTP1D immune complexes derived from EGF-treated cells demonstrated a ligand-dependent coupling between the phosphatase and GRB2 in vivo. Probing of lysates from these cells with glutathione S-transferase (GST) fusion proteins corresponding to the individual domains of GRB2 revealed that this interaction was mediated exclusively by the COOH-terminal SH3 domain of GRB2. Importantly, a GST fusion protein containing the PTP1D SH2 domains was not capable of generating the EGF-induced linkage to GRB2. Additional experiments indicated that neither the binding of the nucleotide exchange factor Sos to GRB2 nor tyrosine phosphorylation of PTP1D was required for EGF-stimulated coupling of PTP1D to GRB2. This is the first demonstration of a growth factor- or cytokine-induced coupling of a protein through an SH3 domain and suggests that GRB2 functions to target PTP1D, in addition to Sos, to the plasma membrane in response to EGF.


Development of Prediction Models for Sickness Absence Due to Mental Disorders in the General Working Population.
Purpose This study investigated if and how occupational health survey variables can be used to identify workers at risk of long-term sickness absence (LTSA) due to mental disorders. Methods Cohort study including 53,833 non-sicklisted participants in occupational health surveys between 2010 and 2013. Twenty-seven survey variables were included in a backward stepwise logistic regression analysis with mental LTSA at 1-year follow-up as outcome variable. The same variables were also used for decision tree analysis. Discrimination between participants with and without mental LTSA during follow-up was investigated by using the area under the receiver operating characteristic curve (AUC); the AUC was internally validated in 100 bootstrap samples. Results 30,857 (57%) participants had complete data for analysis; 450 (1.5%) participants had mental LTSA during follow-up. Discrimination by an 11-predictor logistic regression model (gender, marital status, economic sector, years employed at the company, role clarity, cognitive demands, learning opportunities, co-worker support, social support from family/friends, work satisfaction, and distress) was AUC = 0.713 (95% CI 0.692-0.732). A 3-node decision tree (distress, gender, work satisfaction, and work pace) also discriminated between participants with and without mental LTSA at follow-up (AUC = 0.709; 95% CI 0.615-0.804). Conclusions An 11-predictor regression model and a 3-node decision tree equally well identified workers at risk of mental LTSA. The decision tree provides better insight into the mental LTSA risk groups and is easier to use in occupational health care practice.


Rule-Based Simulation of Multi-Cellular Biological Systems-A Review of Modeling Techniques.
Emergent behaviors of multi-cellular biological systems (MCBS) result from the behaviors of each individual cells and their interactions with other cells and with the environment. Modeling MCBS requires incorporating these complex interactions among the individual cells and the environment. Modeling approaches for MCBS can be grouped into two categories: continuum models and cell-based models. Continuum models usually take the form of partial differential equations, and the model equations provide insight into the relationship among the components in the system. Cell-based models simulate each individual cell behavior and interactions among them enabling the observation of the emergent system behavior. This review focuses on the cell-based models of MCBS, and especially, the technical aspect of the rule-based simulation method for MCBS is reviewed. How to implement the cell behaviors and the interactions with other cells and with the environment into the computational domain is discussed. The cell behaviors reviewed in this paper are division, migration, apoptosis/necrosis, and differentiation. The environmental factors such as extracellular matrix, chemicals, microvasculature, and forces are also discussed. Application examples of these cell behaviors and interactions are presented.


Simulation: Introduction to Correctional Nursing in a Prison Setting.
Simulation can be used to introduce students to critical nursing skills that reduce errors, enhance nurse-patient communication, and improve safety and security when caring for patients in the correctional setting. The interactive environment creates scenarios in a realistic fashion that include experiences that nursing students may encounter in a correctional facility setting, such as aggressive clients, gang-related activities, and noncompliant personalities. Orientation prior to entering the facilities is the key to safety, reducing student anxiety, and enhancing the student's ability to learn. The simulation scenario designed for the correctional setting provides an environment for the teaching and learning of correctional nursing skills and multidisciplinary collaboration. Scenarios are embedded with communication, safety, delegation, and critical thinking education goals. Student evaluations are assessed and reported.


The Presence of Acinetobacter baumannii DNA on the Skin of Homeless People and Its Relationship With Body Lice Infestation. Preliminary Results.
The presence of Acinetobacter baumannii was demonstrated in body lice, however, little is known about the mechanism of natural lice infection. In 2013 and 2014, cross-sectional one-day studies were therefore performed within two Marseille homeless shelters to assess the presence of A. baumannii DNA on human skin, blood and in body lice collected from the same homeless individuals. All 332 participants completed questionnaires, were examined for dermatologic signs, and provided four skin samples (hair, neck, armpits, and pelvic belt), blood samples and body lice (if any). We developed a new real-time PCR tool targeting the ompA/motB gene for the detection of A. baumannii for all collected samples. Blood culture was also performed. Body lice were found in 24/325 (7.4%) of subjects. We showed a prevalence of A. baumannii DNA skin-carriage in 33/305 (10.8%) of subjects. No difference was found in A. baumannii DNA prevalence according to body sites. A strong association between body lice infestation (OR = 3.07, p = 0.029) and A. baumannii DNA skin-carriage was noted. In lice, A. baumannii DNA was detected in 59/219 arthropods (26.9%). All blood cultures and real-time PCR on blood samples were negative for A. baumannii. Lice probably get infected with A. baumannii while biting through the colonized skin and likely transmit the bacteria in their feces. We found no evidence that lice facilitate the invasion of A. baumannii into the blood stream. Further investigations are needed to compare phenotypic and genotypic features of A. baumannii isolates from human skin and lice from the same individuals.


Evaluation of genotoxicity of Pseudevernia furfuracea (L.) Zopf by RAPD analysis.
We investigated the suitability and applicability of Pseudevernia furfuracea (L.) Zopf for environmental genotoxicity assessment. P. furfuracea lichen specimens were collected from 10 different Pinus species, in every 5 km, starting from around an iron-steel factory located in the central area of Karabük Province up to Yenice Forest. The impact of the pollution sources such as iron-steel factory, roads and railroads, industry, heavy traffic, and waste treatment plants on the heavy metal accumulation in lichens is known. DNA changes in P. furfuracea samples exposed naturally to various polluted sites were analyzed by RAPD to know the influence of the environmental pollution on the hereditary material of the organisms. Twenty-five different primers were tested and 10 yielded clear and reproducible bands. The present study shows the suitability of the lichen samples for the detection of genotoxicity and also provides information about the level of potential genotoxic agents around a steel mill.


Assessment of Midline Lingual Point-Pressure Somatosensation Using Von Frey Hair Monofilaments.
Detection and discrimination threshold estimates for oral point pressure are assessed using Von Frey Hair monofilaments. Consistent with previously published protocols, threshold estimates are determined using a two-interval forced choice (2-IFC) paradigm with a three down/one up approach. Detection threshold estimates determine the mean force in which a participant can identify the presence of pressure. During the detection threshold procedure, the participant is instructed to choose which of two sequentially presented observation intervals contained the tactile test stimulus. If the participant performs three correct detections in a row (i.e., 3 'hits'), the researcher decreases the stimulus to the next lower target force level. With one incorrect detection (a 'miss'), the researcher increases the force delivered to the next higher level. This threshold estimation approach is known as a 3-down/1-up adaptive staircase. Reponses are recorded on a paper ballot, and a participant's estimated threshold is defined as the geometric mean of five reversals. During the discrimination threshold procedure, the participant is asked to make a choice between two serially presented stimuli as to which is the "harder" or "stronger" pressure. The same scoring of 'hits', 'misses', and stopping points are used. Detection and discrimination testing for oral point pressure at tongue midline takes approximately 20 min to complete. Using these commercially-available clinical tools, individual touch sensation profiles for the midline tongue can be achieved in a relatively time and cost effective means.


Identification and characterization of prostein, a novel prostate-specific protein.
In this report, we describe the application of a systematic, genome-based approach to identify prostein, a novel prostate-specific protein expressed in normal and malignant prostate tissues. Characterization of the prostein gene shows that prostein cDNA encodes a 553-amino acid protein. The protein is predicted to be a type IIIa plasma membrane protein with a cleavable signal peptide and 11 transmembrane-spanning regions. The prostein gene is located on chromosome 1 at the WI-9641 locus between q32 and q42. Prostein mRNA is shown to be uniquely expressed in normal and cancerous prostate tissues using Northern blot, eDNA microarray, and real-time PCR analyses. Furthermore, prostein mRNA expression does not appear to be prostate tumor grade related and is restricted exclusively to prostate cell lines. Immunohistochemical staining using a mouse monoclonal antibody generated against prostein demonstrates that this protein is specifically detected in prostate tissues both at the plasma membrane and in the cytoplasm. Prostein expression is androgen responsive because treatment of LNCaP cells with androgen up-regulates prostein message and protein expression levels. These results validate prostein as a prostate-specific marker with potential utility in the diagnosis and treatment of prostate cancer.


High triglyceride-glucose index at admission is a predictor of post-stroke cognitive impairment in patients with acute ischemic stroke.
Post-stroke cognitive impairment (PSCI) is a very common complication of ischemic stroke (IS). Triglyceride-glucose (TyG) index was an effective alternative marker of insulin resistance (IR). This prospective study was designed to explore the correlation between TyG index and PSCI. Between January 1 2021 to June 30 2022, consecutive patients with first onset IS were enrolled prospectively. Baseline information was collected at admission and fasting blood was drawn the next morning. Montreal Cognitive Assessment (MoCA) was used to evaluate cognitive function at three months after stroke. Multiple regression analysis was used to explore the correlation between PSCI and TyG. Receiver operating characteristic (ROC) was performed to evaluate the predictive ability. Ultimately, 313 patients were enrolled in this study. The TyG index was higher in patients with PSCI than those without PSCI (8.99 (8.55, 9.54) vs. 8.61(8.25, 8.87), P<0.001). The spearman correlation analysis indicated that TyG index was negatively correlated with MoCA score (r=-0.272, P<0.001). The multivariate logistic regression analysis demonstrated that TyG index was correlated with PSCI independently (P<0.001) regardless of whether the patients had diabetes or not. The area under curve (AUC) of the ROC was 0.684 (95%CI=0.635-0.768, P<0.001). The optimal cutoff value of TyG index for predicting PSCI was 8.81, with a sensitivity of 61.7% and a specificity of 73.6%. A higher TyG index level at admission was independently correlated with increased risk of PSCI three months later and could be used as a predictor.


P-wave dispersion and its relationship to aortic stiffness in patients with acute myocardial infarction after cardiac rehabilitation.
The aim of our study was to investigate the P-wave dispersion from standard electrocardiograms (ECGs) in patients with acute myocardial infarction (AMI) after cardiac rehabilitation (CR) and determine its relation to arterial stiffness. This is a prospective study included 33 patients with AMI and successfully re-vascularized by percutaneous coronary intervention (PCI) underwent CR. Left ventricular ejection fraction (LVEF) was measured by biplane Simpson's method. Left atrium (LA) volume was calculated. The maximum and minimum durations of P-waves (Pmax and Pmin, respectively) were detected, and the difference between Pmax and Pmin was defined as P-wave dispersion (Pd = Pmax-Pmin). Aortic elasticity parameters were measured. LVEF was better after CR. The systolic and diastolic blood pressures decreased after CR, these differences were statistically significant. With exercise training, LA volume decreased significantly. Pmax and Pd values were significantly shorter after the CR program. The maximum and minimum P-waves and P-wave dispersion after CR were 97 ± 6 ms, 53 ± 5 ms, and 44 ± 5 ms, respectively. Aortic strain and distensibility increased and aortic stiffness index was decreased significantly. Aortic stiffness index was 0.4 ± 0.2 versus 0.3 ± 0.2, P = 0.001. Aortic stiffness and left atrial volume showed a moderate positive correlation with P-wave dispersion (r = 0.52, P = 0.005; r = 0.64, P < 0.001, respectively). This study showed decreased arterial stiffness indexes in AMI patient's participated CR, with a significant relationship between the electromechanical properties of the LA that may raise a question of the preventive effect of CR from atrial fibrillation and stroke in patients with acute myocardial infarction.


Using the Cancer Risk Management Model to evaluate colorectal cancer screening options for Canada.
Several screening methods for colorectal cancer (crc) are available, and some have been shown by randomized trials to be effective. In the present study, we used a well-developed population health simulation model to compare the risks and benefits of a variety of screening scenarios. Tests considered were the fecal occult blood test (fobt), the fecal immunochemical test (fit), flexible sigmoidoscopy, and colonoscopy. Outcomes considered included years of life gained, crc cases and deaths prevented, and direct health system costs. A natural history model of crc was implemented and calibrated to specified targets within the framework of the Cancer Risk Management Model (crmm) from the Canadian Partnership Against Cancer. The crmm-crc permits users to enter their own parameter values or to use program-specified base values. For each of 23 screening scenarios, we used the crmm-crc to run 10 million replicate simulations. Using base parameter values and some user-specified values in the crmm-crc, and comparing our screening scenarios with no screening, all screening scenarios were found to reduce the incidence of and mortality from crc. The fobt was the least effective test; it was not associated with lower net cost. Colonoscopy screening was the most effective test; it had net costs comparable to those for several other strategies considered, but required more than 3 times the colonoscopy resources needed by other approaches. After colonoscopy, strategies based on the fit were predicted to be the most effective. In sensitivity analyses performed for the fobt and fit screening strategies, fobt parameter values associated with high-sensitivity formulations were associated with a substantial increase in test effectiveness. The fit was more cost-effective at the 50 ng/mL threshold than at the 100 ng/mL threshold. The crmm-crc provides a sophisticated and flexible environment in which to evaluate crc control options. All screening scenarios considered in this study effectively reduced crc mortality, although sensitivity analyses demonstrated some uncertainty in the magnitude of the improvements. Where possible, local data should be used to reduce uncertainty in the parameters.


A morbilliform variant of vancomycin-induced linear IgA bullous dermatosis.
Linear IgA bullous dermatosis is an autoimmune blistering disease characterized clinically by the presence of small tense blisters and immunologically by the presence of IgA at the dermal-epidermal junction. Idiopathic, systemic disease-related, and drug-related versions of this disorder have been described, with the latter most commonly associated with vancomycin. We describe 2 patients with vancomycin-associated linear IgA bullous dermatosis who presented with a morbilliform eruption that lacked blistering. Lesional and perilesional tissue from each patient was examined by light microscopy and direct immunofluorescence. Histopathologic examination findings revealed vacuolar interface dermatitis with a mixed inflammatory infiltrate and occasional eosinophils, consistent with a drug eruption. Direct immunofluorescence revealed IgA deposited in a linear pattern at the dermoepidermal junction. In both patients, the results of indirect immunofluorescence using both IgG and IgA were negative. These cases highlight the existence of a new form of linear IgA bullous dermatosis presenting as a morbilliform drug eruption. Both patients were following extensive medication regimens, including use of multiple antibiotics. The diagnosis of linear IgA bullous dermatosis allowed us to target vancomycin as the likely allergen and begin treatment. In light of these findings, direct immunofluorescence may be a useful diagnostic adjunct in determining the cause of drug eruptions.


Cancer incidence in patients with hereditary hemorrhagic telangiectasia.
Hereditary hemorrhagic telangiectasia (HHT) is a genetic disorder characterized by deficiency in endoglin, an angiogenic protein. We previously showed that HHT, in which systemic endoglin expression is reduced, was associated with better survival outcomes in cancer patients (Duarte et al. in Cancer Epidemiol Biomarkers Prev 23:117-125, 2014). Here, we evaluated whether HHT was associated with reduced cancer incidence. A matched case-control analysis using SEER Medicare was conducted to evaluate the effect of HHT on diagnosis with breast, colorectal, lung, or prostate cancer between 2000 and 2007 (n = 633,162). Cancer and non-cancer patients were matched on age, sex, SEER registry region, and length of the ascertainment period for HHT. We assessed crude association using a McNemar's test and then adjusted for demographic variables, cancer type, cancer stage, comorbidities, and ascertainment period with a conditional logistic regression model for cancer incidence. The McNemar's test showed no significant association between HHT and cancer incidence (p = 0.74). Adjusting for covariates with the conditional logistic regression model did not change the result [HHT odds ratio 0.978; 95 % CI (0.795, 1.204)]. The lack of association between HHT and cancer incidence is unexpected given the previously discovered significant association between HHT and improved survival outcomes (Duarte et al. in Cancer Epidemiol Biomarkers Prev 23:117-125, 2014). We conclude that the protective effect of reduced systemic endoglin expression in cancer is specific to cancer progression through its effect on vascularization and other stromal effects but does not extend to cancer initiation.


Robust Nanoparticles Detection From Noisy Background by Fusing Complementary Image Information.
This paper studies the problem of detecting the presence of nanoparticles in noisy transmission electron microscopic (TEM) images and then fitting each nanoparticle with an elliptic shape model. In order to achieve robustness while handling low contrast and high noise in the TEM images, we propose an approach to fuse two kinds of complementary image information, namely, the pixel intensity and the gradient (the first derivative in intensity). Our approach entails two main steps: 1) the first step is to, after necessary pre-processing, employ both intensity-based information and gradient-based information to process the same TEM image and produce two independent sets of results and 2) the subsequent step is to formulate a binary integer programming (BIP) problem for conflict resolution among the two sets of results. Solving the BIP problem determines the final nanoparticle identification. We apply our method to a set of TEM images taken under different microscopic resolutions and noise levels. The empirical results show the merit of the proposed method. It can process a TEM image of 1024×1024 pixels in a few minutes, and the processed outcomes appear rather robust.


Comparison of Two Schizophyllum commune Strains in Production of Acetylcholinesterase Inhibitors and Antioxidants from Submerged Cultivation.
In recent years, fungi have been recognized as producers of acetylcholinesterase (AChE) inhibitors, agents important for the prevention of Alzheimer's disease (AD). This study aimed to examine the AChE inhibitory, the antioxidative and antibacterial activity of two different Schizophyllum commune strains that originated from Serbia (SRB) and Italy (IT). Submerged cultivation of grown mycelia (M) and fermentation broth (F) of ethanol (EtOH) and polysaccharide (PSH) extracts lasted for 7, 14, 21 and 28 days. For AChE activity Ellman method was performed, while for antioxidative activity, sevendifferent assays were conducted: DPPH, ABTS, FRAP, SOA, OH, NO together with total phenolic content. Antimicrobial screen, LC-MS/MS technique and FTIR measurements were performed. Different isolates exhibited different AChE activity, with PSH being the strongest (SRB, M, 28 days IC90 79.73 ± 26.34 µg/mL), while in EtOH extracts, IT stood out (F, 14 days, IC50 0.8 ± 0.6 µg/mL). PSH extracts (7 days) exhibit significant antioxidative activity (AO), opposite to EtOH extracts where 14 and 21days periods stood out. Only tw extracts showed antibacterial activity. Following LC-MS/MS analysis p-hydroxybenzoic and gallic acids were the most abundant phenolics. PSH extracts demonstrated remarkable results, making this study debut and introducing S. commune as a valuable resource of AChE inhibitors.


Cryptosporidium parvum biliary tract infection in adult immunocompetent and immunosuppressed mice.
Biliary tract infection by Cryptosporidium parvum is a frequent complication of intestinal cryptosporidiosis in immunosuppressed patients. Although biliary tract infection can be produced in immunosuppressed models as a late complication of intestinal infection, there is no infection model in immunocompetent animals. A murine model of biliary tract cryptosporidiosis was developed by direct intra-gall bladder injection of C. parvum oocysts. In adult immunocompetent mice, intracellular parasitic stages were detected in the epithelium of the common bile duct in all animals on day 7 post-inoculation (p.i.). These findings were associated with a strong inflammatory reaction. Infection was cleared between days 14 and 21 p.i. All animals developed significant levels of specific serum and bile IgG, IgA and IgM. Dexamethasone treatment resulted in the inability of animals to eradicate the parasite and the establishment of ileal parasitism. This model can be used to study the immunological mechanisms involved in the control of biliary cryptosporidiosis.


Functional domains of the human estrogen receptor.
Two domains of the human estrogen receptor, responsible for hormone binding (region E) and tight nuclear binding (region C), are essential for the receptor to activate efficiently the transcription of estrogen-responsive genes. Region D, which joins the DNA- and hormone-binding domains, can be altered without affecting activation. Deletion of the N-terminal domain (region A/B) has no effect on activation of a reporter gene containing a vitellogenin estrogen-responsive element (ERE) and the HSV-tk promoter, whereas it severely impairs activation of the human pS2 gene promoter. Deletion of most or all of the hormone-binding domain leads to only about 5% constitutive transcriptional activity, yet these mutants appear to bind efficiently to an ERE in vivo. Apparently, region C recognizes the ERE of target genes, and the hormone-binding domain plays an essential role for efficient activation of transcription.


Phthalate Exposures and MicroRNA Expression in Uterine Fibroids: The FORGE Study.
Phthalates are associated with multiple, adverse reproductive outcomes including increased risk of uterine leiomyoma (fibroids). Phthalates can interact with epigenetic modifications including microRNAs (miRNAs), which help regulate processes crucial to fibroid pathogenesis. However, no prior study has examined the influence of phthalates on miRNA expression in fibroid tumors. We conducted a preliminary, cross-sectional study to examine the associations between phthalate exposures and miRNA expression levels in fibroid tumors and to explore potential effect modification by race/ethnicity. We quantified expression levels of 754 miRNAs in fibroid tumor samples and analyzed spot urine samples for phthalate metabolites collected from 45 pre-menopausal women undergoing surgery for fibroid treatment at an academic hospital. Associations between miRNA levels in fibroids and phthalate biomarkers were evaluated using linear regression adjusting for age, race/ethnicity, and body mass index (BMI). Statistical tests were adjusted for multiple comparisons. We also performed in silico Ingenuity Pathway Analysis to identify the biological pathways that are regulated by phthalate-associated miRNAs. Mono-hydroxybutyl phthalate and mono(2-ethyl-5-hydroxyhexyl) phthalate were positively associated with miR-10a-5p (β = 0.76, 95% CI = [0.40, 1.11]) and miR-577 (β = 1.06, 95% CI = [0.53, 1.59]), respectively. A total of 8 phthalate-miRNA associations varied by race/ethnicity (qinteraction < 0.10). Pathway analysis revealed that mRNA gene targets of phthalate-associated miRNAs were significantly associated with multiple fibroid-related processes including angiogenesis, apoptosis, and proliferation of connective tissues. Collectively, these data suggest that exposures to some phthalates are associated with miRNA in fibroids, and that associations may vary by race/ethnicity. Validation of these findings may provide insight into mechanisms underlying associations between phthalates and fibroids and contribute to novel hypotheses regarding racial/ethnic disparities in fibroids.


Lipopolysaccharide Impedes Bone Repair in FcγRIIB-Deficient Mice.
Chronic inflammation contributes to the development of skeletal disorders in patients with systemic lupus erythematosus (SLE). Activation of the host immune response stimulates osteoclast activity, which in turn leads to bone loss. Regenerating bone in the inflammatory microenvironments of SLE patients with critical bone defects remains a great challenge. In this study, we utilized lipopolysaccharide (LPS) to imitate locally and systemically pathogenic bacterial infection and examined the bone regeneration performance of LPS-associated mandibular and tibial bone regeneration impairment in FcγRIIB-/- mice. Our results indicated that a loss of FcγRIIB alleviates bone regeneration in both mandibles and tibiae. After LPS induction, FcγRIIB-/- mice were susceptible to impaired fracture healing in tibial and mandibular bones. LPS decreased the mineralization to collagen ratio in FcγRIIB-/- mice, indicating a mineralization defect during bone repair. An osteoblast-associated gene (Col1a1) was attenuated in FcγRIIB-deficient mice, whereas Bglap, Hhip, and Creb5 were further downregulated with LPS treatment in FcγRIIB-/- mice compared to FcγRIIB-/- mice. Alpl and Bglap expression was dcreased in osteoblasts derived from bone chips. An osteoclast-associated gene, Tnfsf11/Tnfrsf11 ratio, ewas increased in LPS-induced FcγRIIB-/- mice and in vitro. Furthermore, systemic LPS was relatively potent in stimulating production of pro-inflammatory cytokines including TNF-α, IL-6, and MCP-1 in FcγRIIB-/- mice compared to FcγRIIB-/- mice. The levels of TNF-α, IFN-β, IL-1α, and IL-17A were increased, whereas IL-10 and IL-23 were decreased in FcγRIIB-/- mice treated locally with LPS. These findings suggest that both local and systemic LPS burden can exacerbate bone regeneration impairment, delay mineralization and skeletal repair, and induce inflammation in SLE patients.


Titan cell production in Cryptococcus neoformans reshapes the cell wall and capsule composition during infection.
Cryptococcus neoformans is a human fungal pathogen that often causes infections in immunocompromised individuals. Upon inhalation into the lungs C. neoformans differentiates into cells with altered size and morphology, including production of large titan cells. Titan cells possess thickened cell wall and dense, cross-linked capsule when compared to in vitro grown cells. In addition, titan cells have increased cell wall chitin that is associated with a detrimental anti-inflammatory immune response. Here we examined the cell wall and capsule composition of in vitro, in vivo typical-sized and in vivo titan cells using High Performance Liquid Chromatography (HPLC). The monomer composition of cell wall polysaccharides showed that in vivo C. neoformans cells contained more glucosamine and less glucose than in vitro cells, suggesting alteration in abundance of both chitin and glucans, respectively. Low levels of galactosamine were also detected in carbohydrates from both in vivo and vitro cells. Within the in vivo cell population, differences in the proportions of cell wall and capsule monomers between typical and titan cells were also observed. Taken together, these results demonstrate that C. neoformans reshapes its cell wall and capsule composition during infection. These cell wall and capsule alterations likely help C. neoformans escape recognition by, and allow modulation of, the host immune system.


Usefulness of time-resolved MR angiography in spinal dural arteriovenous fistula (SDAVF)-a systematic review and meta-analysis.
Spinal dural arteriovenous fistulas (SDAVFs) constitute the most common type of spinal vascular malformations. Their diagnosis requires spinal digital subtraction angiography (DSA), which is time-consuming, requires catheterizing many vessels, and exposes patient to a high radiation and contrast doses. This study aims to evaluate the usefulness of time-resolved MR angiography (TR-MRA) in SDAVF diagnosis. We performed a systematic review of the PubMed and EMBASE databases followed by a meta-analysis. TR-MRA was an index test, and spinal DSA was a reference. Of the initial 324 records, we included 4 studies describing 71 patients with SDAVFs. In 42 cases, TR-MRA was true positive, and in 21 cases, it was true negative. We found 7 false-positive cases and 1 false negative. TR-MRA allowed for shunt level identification in 39 cases. Of these, the predicted level was correct in 23 cases (59%), to within 1 level in 38 cases (97.4%) and to within 2 levels in 39 cases (100%). The diagnostic odds ratio was 72.73 (95% CI [10.30; 513.35]), z = 4.30, p value < 0.0001. The pooled sensitivity was 0.98 (95% CI [0.64; 1.00]), and the pooled specificity was 0.79 (95% CI [0.10; 0.99]). The AUC of the SROC curve was 0.9. TR-MRA may serve as a preliminary study to detect SDAVFs and localize the shunt level with sensitivity and specificity as high as 98% and 79%, respectively. Unless the TR-MRA result is unequivocal, it should be followed by a limited spinal DSA.


A systemic lymphoproliferative disorder with morphologic features of Castleman's disease. Pathological findings in 15 patients.
This report describes the nodal and extranodal lesions observed in 15 patients with a generalized disorder that had been histologically diagnosed as Castleman's disease. The disorder was characterized by severe constitutional symptoms, constant involvement of multiple peripheral lymph nodes, and frequent hepatosplenomegaly, in association with clinical and laboratory features reminiscent of a "collagen disease." The clinical course was chronic, with remissions and exacerbations in seven patients, and aggressive and fatal in eight. The material examined included multiple lymph node biopsies, four surgical specimens of spleen, one open lung biopsy, and material from four autopsies. The diagnostic morphological findings were observed in the nodes and were represented by the following histologic triad: diffuse marked plasmacytosis, from the medulla to the subcapsular areas; prominence of the germinal centers; and good preservation of the architecture. One variant of this basic pattern featured abundant immunoblasts and blood vessels. The process appears to be a systemic reactive proliferation of B-lymphocytes, perhaps resulting from faulty immune regulation. Morphologic similarities indicate a relationship between this multicentric disorder and Castleman's disease of plasmacellular type. However, there are distinct differences between them in clinical presentation and evolution, and, consequently, in therapeutic approach.


Effects of Cover Crop Species and Season on Population Dynamics of Escherichia coli and Listeria innocua in Soil.
Cover crops provide several ecosystem services, but their impact on enteric bacterial survival remains unexplored. The influence of cover cropping on foodborne pathogen indicator bacteria was assessed in five cover crop/green manure systems: cereal rye, hairy vetch, crimson clover, hairy vetch-rye and crimson clover-rye mixtures, and bare ground. Cover crop plots were inoculated with Escherichia coli and Listeria innocua in the fall of 2013 and 2014 and tilled into the soil in the spring to form green manure. Soil samples were collected and the bacteria enumerated. Time was a factor for all bacterial populations studied in all fields (P < 0.001). E. coli levels declined when soil temperatures dipped to <5°C and were detected only sporadically the following spring. L. innocua diminished somewhat but persisted, independently of season. In an organic field, the cover crop was a factor for E. coli in year 1 (P = 0.004) and for L. innocua in year 2 (P = 0.011). In year 1, E. coli levels were highest in the rye and hairy vetch-rye plots. In year 2, L. innocua levels were higher in hairy vetch-rye (P = 0.01) and hairy vetch (P = 0.03) plots than in the rye plot. Bacterial populations grew (P < 0.05) or remained the same 4 weeks after green manure incorporation, although initial reductions in L. innocua numbers were observed after tilling (P < 0.05). Green manure type was a factor only for L. innocua abundance in a transitional field (P < 0.05). Overall, the impacts of cover crops/green manures on bacterial population dynamics in soil varied, being influenced by bacterial species, time from inoculation, soil temperature, rainfall, and tillage; this reveals the need for long-term studies.


Image-guided combination chemotherapy and photodynamic therapy using a mitochondria-targeted molecular probe with aggregation-induced emission characteristics.
Subcellular targeted cancer therapy and in situ monitoring of therapeutic effect are highly desirable for clinical applications. Herein, we report a series of probes by conjugating zero (TPECM-2Br), one (TPECM-1TPP) and two (TPECM-2TPP) triphenylphosphine (TPP) ligands to a fluorogen with aggregation-induced emission (AIE) characteristics. The probes are almost non-emissive as molecularly dissolved species, but they can light up in cell cytoplasm or mitochondria. TPECM-2TPP is found to be able to target mitochondria, depolarize mitochondria membrane potential and selectively exert potent chemo-cytotoxicity on cancer cells. Furthermore, it can efficiently generate singlet oxygen with strong photo-toxicity upon light illumination, which further enhances its anti-cancer effect. On the other hand, TPECM-1TPP can also target mitochondria and generate singlet oxygen to trigger cancer cell apoptosis, but it shows low cytotoxicity in dark. Meanwhile, TPECM-1TPP can report the cellular oxidative stress by visualizing the morphological changes of mitochondria. However, TPECM-2Br does not target mitochondria and shows no obvious anticancer effect either in dark or under light illumination. This study thus highlights the importance of molecular probe design, which yields a new generation of subcellular targeted molecular theranostic agents with multi-function, such as cancer cell imaging, chemotherapy, photodynamic therapy, and in situ monitoring of the therapeutic effect in one go.