Airway Obstruction from a Tracheal Mass in a 5-Year-Old: A Case Report.
Stridor is a common presenting symptom for children in emergency departments (EDs) and usually represents an infectious process, such as croup, or aspiration of a foreign body. We present the case of an otherwise healthy 5-year-old girl with episodic increased work of breathing for several months that was initially diagnosed as asthma by her primary care physician. She subsequently presented to the ED with acutely worsening noisy breathing and dyspnea. Patient and parent denied any recent foreign body ingestions or choking episodes. We gave multiple doses of racemic epinephrine in the ED without symptom improvement. A lateral neck x-ray showed an occlusive subglottic airway mass. Otolaryngology (ENT) evaluation demonstrated an 85% airway occlusion. The mass was partially resected, resolving all of her respiratory symptoms. Although primary airway tumors in children are rare, they must be considered on the differential diagnosis of new noisy breathing or respiratory distress. Failure to diagnose these tumors in a timely manner can be life-threatening.


PD-1/PD-L1 Inhibitors in Patients With Preexisting Autoimmune Diseases.
Autoimmune diseases and malignant tumors are the two hotspots and difficulties that are currently being studied and concerned by the medical field. The use of PD-1/PD-L1 inhibitors improves the prognosis of advanced tumors, but excessive immune responses can also induce immune-related adverse events (irAEs). Due to this concern, many clinical trials exclude cancer patients with preexisting autoimmune disease (AID). This review outlines the possible mechanisms of irAE, discusses the safety and efficacy of PD-1/PD-L1 inhibitors in cancer patients with preexisting AID, and emphasizes the importance of early recognition, continuous monitoring, and multidisciplinary cooperation in the prevention and management of cancer patients with preexisting AID.


Targeting TDO in cancer immunotherapy.
Tryptophan-2,3-dioxygenase (TDO) is a homotetrameric heme-containing protein catalyzing the initial step in the kynurenine pathway, which oxidates the 2,3-double bond of the indole ring in L-tryptophan and catalyzes it into kynurenine (KYN). The upregulation of TDO results in a decrease in tryptophan and the accumulation of KYN and its metabolites. These metabolites can affect the proliferation of T cells. Increasing evidence demonstrates that TDO is a promising therapeutic target in the anti-tumor process. Despite its growing popularity, there are only a few reviews focusing on TDO in tumors. Hence, we herein review the biological features and regulatory mechanisms of TDO. Additionally, we focus on the role of TDO in the anti-tumor immune response in different tumors. Finally, we also provide our viewpoint regarding the future developmental directions of TDO in cancer research, especially in relation to the development and application of TDO inhibitors as novel cancer treatments.


The veno-venous bypass in liver transplantation: an unfinished product.
Veno-venous bypass (VVB) using a patent para-umbilical vein during liver transplantation (LT) has not been reported previously. Here, we report the decompression of the porto-mesenteric compartment via a patent para-umbilical vein in a patient needing a VVB during LT. Pre-transplant CT-scan showed a large patent para-umbilical vein. A femoro-axillary percutaneous VVB was installed prior to abdominal opening to decompress massive collateral veins in the abdominal wall. The para-umbilical vein was stapled and its proximal end was cannulated and connected to the VVB. The severe atrophy of the native liver allowed to place the whole liver graft in the "liver fossa" while maintaining the native liver pulled toward the left side and connected to the VVB. This maneuver maintained splanchnic venous decompression during latero-lateral cavo-caval anastomosis. The "portal" cannula was clamped and removed. The native portal vein was clamped and divided. Standard portal, arterial, and biliary reconstructions were then performed. A patent para-umbilical vein, present in one third of patients with portal hypertension could be used whenever VVB is indicated during LT in this setting.


[Electroencephalographic correlates of schizophrenic type mental disorders at an early age in childhood].
Electroencephalographic study was performed in 30 children of 1-3 years old from the group with the high risk of schizophrenia. Clinical observation of the patients was performed in the period of EEG recording and follow-up study was also made during 10-12 years. Three groups of patients were picked out with the differences in both clinical and electrophysiologic indices. Bundle beta-activity was registered on EEG in the cases of an active schizophrenic process. A presence of the spindles of the sleep and their dominance on EEG were characteristic for children with nonprocessual disorders of psychopathic-like type with disinhibition of the drives. Hypersynchronism of delta- and theta-activities were observed in the cases of schizotypic diathesis and an early children's autism with the predominance of paroxysmal somato-autonomic disorders.


Molecular analyses on Neospora caninum-triggered NETosis in the caprine system.
Neospora caninum is an obligate intracellular protozoan parasite causing serious reproductive disorders in large and small ruminants worldwide. Polymorphonuclear neutrophils (PMN) react against multiple invading pathogens through different mechanisms including the release of neutrophil extracellular traps (NETs). Here, in vitro interactions of caprine PMN and N. caninum tachyzoites were studied. Scanning electron microscopic- and immunofluorescence-analyses demonstrated that caprine PMN undergo NETosis upon contact with tachyzoites of N. caninum, extruding filaments that entrap parasites. Detailed co-localization studies of N. caninum tachyzoite-induced NETs revealed the presence of PMN-derived DNA being decorated with histones (H1, H2A/H2B, H3,H4) and neutrophil elastase (NE) corroborating the molecular characteristics of classical mammalian NETs. As a new result for parasite-induced NETosis, we identified pentraxin and cathepsin B in N. caninum-triggered NETs. Nonetheless, functional inhibition assays revealed that during caprine NET formation triggered by N. caninum different molecular signaling pathways are induced, when compared to other apicomplexan parasites or host species. As such, N. caninum-induced NETosis appears to be influenced by MPO but independent of NADPH oxidase, SOCE, ERK1/2 and p38 MAPK activities. Furthermore, the inhibition of PMN autophagy via blockage of the PI3K-mediated signaling pathway failed to influence tachyzoite-induced NETosis. Since N. caninum-tachyzoites induced caprine NETosis, this effector mechanism should be considered as an early host immune response during acute caprine neosporosis.


Vigilant keratinocytes trigger pathogen-associated molecular pattern signaling in response to streptococcal M1 protein.
The human skin exerts many functions in order to maintain its barrier integrity and protect the host from invading microorganisms. One such pathogen is Streptococcus pyogenes, which can cause a variety of superficial skin wounds that may eventually progress into invasive deep soft tissue infections. Here we show that keratinocytes recognize soluble M1 protein, a streptococcal virulence factor, as a pathogen-associated molecular pattern to release alarming inflammatory responses. We found that this interaction initiates an inflammatory intracellular signaling cascade involving the activation of the mitogen-activated protein kinases extracellular signal-regulated kinase (ERK), p38, and Jun N-terminal protein kinase and the subsequent induction and mobilization of the transcription factors NF-κB and AP-1. We also determined the imprint of the inflammatory mediators released, such as interleukin-8 (IL-8), growth-related oncogene alpha, migration inhibitory factor, extracellular matrix metalloproteinase inducer, IL-1α, IL-1 receptor a, and ST2, in response to streptococcal M1 protein. The expression of IL-8 is dependent on Toll-like receptor 2 activity and subsequent activation of the mitogen-activated protein kinases ERK and p38. Notably, this signaling seems to be distinct for IL-8 release, and it is not shared with the other inflammatory mediators. We conclude that keratinocytes participate in a proinflammatory manner in streptococcal pattern recognition and that expression of the chemoattractant IL-8 by keratinocytes constitutes an important protective mechanism against streptococcal M1 protein.


Selective eaters: a retrospective case note study.
The literature on children who present with selective eating is limited to single case studies. This study aims to provide a descriptive clinical profile for these children. Case notes of 33 patients between 4 and 14 years of age, seen in two clinics over a 4-year-period were studied. Over two-thirds of the cases were boys. A significant minority had poor growth or weight gain. The children suffered from anxiety, obsessive compulsive symptoms, both food and non-food related and often had social and school difficulties. Mealtimes caused immense anxiety for family members and frequent mealtime battles was reported by the parents of the younger selective eaters, whereas parents of older children seemed to have given up trying to change their child's eating habits. A history of depression in at least one parent was found in a third of the parental couples.


Comparison of 3,4-dimethoxyphenylethylamine treated plasma from chronic schizophrenics and controls.
Plasma samples were collected from chronic schizophrenic patients not on drug therapy and from non-psychotic hospital volunteet controls and incubated with 3,4-dimethoxywhenylethylamine (DMPEA) when injected into monoamine oxidase inhibitor (MAOI) pretreated mice who were then aggregated (groups of 5 ea.) for four hours, the two kinds of plasma produced results so similar that no differences between them could be observed. Doubling the amount of plasma and DMPEA likewise failed to differentiate plasma origin. In our hands no differentiation between plasma samples from control subjects and schizophrenic patients could be determined under the conditions employed.


The Charnley versus the Christiansen total hip arthroplasty. A comparative clinical study.
Charnley total hip arthroplasties (THA) and Christiansen THA were carried out in two concurrent groups of patients, consisting of 113 hips in 87 patients and 90 hips in 81 patients respectively. The mean age of the patients was about 65 years, two thirds had idiopathic arthrosis, and only two had rheumatic hip disease. The two groups of patients were similar with regard to sex, age, weight, etiology of arthrosis, and standard laboratory tests, but different with regard to the surgeons who operated on them. Postoperative films showed that the stems of the prostheses were in the neutral and the varus position in the Christiansen group, and in the valgus, neutral and varus position in the Charnley group. Neither group had any deaths, or deep infections in the postoperative period. Only minor postoperative complications were noted. At follow-up, 5-8 years later, the failure (revision) rate of Charnley THA to Christiansen THA was 1 to 7.7 and 1 Charnley cup was revised as against 19 Christiansen cups. The methodological weaknesses in the comparative clinical trial did not explain the inferior long-term results of the Christiansen THA.


Thermodynamic Ultrastability of a Polymer Glass Confined at the Micrometer Length Scale.
We employ fast scanning calorimetry to assess the thermodynamic state attained after a given cooling rate and the molecular mobility of glassy poly(4-tert-butylstyrene) confined at the micrometer length scale. We show that, for such a large confinement length scale, thermodynamic states with a fictive temperature (T_{f}) 80 K below the polymer glass transition temperature (T_{g}) are attained, which allows to bypass the geological timescales required for bulk glasses. Access to such states is promoted by a fast mechanism of equilibration. Importantly, the tremendous T_{f} decrease takes place while the molecular mobility remains bulklike, indicating marked decoupling between vitrification kinetics and molecular mobility.


Dual energy X-ray absorptiometry outcomes in male COPD patients after treatment with different glucocorticoid regimens.
To compare bone mineral density (BMD) outcomes of patients who received continuous oral systemic glucocorticoids (GCs) with BMD outcomes of patients who received multiple GC courses, oral or IV. Cross-sectional study. Eighty-six white men with COPD selected from the outpatient clinic for pulmonary diseases. Data analysis from medical records, bone densitometry, and pulmonary function tests of consecutive selected patients. Inclusion period into the study was exactly 1 year. Ten patients received oral prednisolone daily (group 1). Eleven patients were treated for several exacerbations with multiple systemic prednisolone courses, up to a period of 2 weeks per course, with a cumulative dose of > or = 1,000 mg (group 2). Likewise, 28 patients were treated with multiple systemic prednisolone courses, but with a cumulative dose < 1,000 mg (group 3). Thirty-seven patients were never treated with systemic prednisolone, and partly with inhaled corticosteroids (ICS) [group 4]. All groups were balanced for age and pack-years of smoking. In group 2, body mass index (BMI) and FEV(1) were lowest and hyperinflation was highest. The cumulative systemic prednisolone dose was highest in group 1, irrespective of the additional ICS treatments. Dual energy x-ray absorptiometry scanning of the lumbar spine, total hip, and femoral neck regions revealed a T score < or = 2.5 SD in 27 patients (31%), 31 patients (36%), and 34 patients (40%), respectively. BMD outcomes at any site were lower in patients receiving multiple systemic prednisolone courses > 1,000 mg, cumulatively (group 2), compared to the other groups, and these values were (mean +/- 1 SD) 0.759 +/- 0.238 g/cm(2), 0.683 +/- 0.115 g/cm(2), and 0.686 +/- 0.125 g/cm(2), respectively (p < 0.0001). Multivariate regression analysis revealed a correlation between the cumulative dose of prednisolone in group 2 and BMD of the lumbar spine (adjusted r = 0.48; p < 0.01). At the total hip and femoral neck regions, only a correlation between BMI and BMD was observed (adjusted r = 0.65 and 0.58, respectively; p < 0.0001 for both sites). Despite a far lower cumulative GC dose in comparison with patients treated with systemic corticosteroids continuously, after adjusting for BMI and lung function, osteoporosis of the lumbar spine was most frequent in patients receiving > 1,000 mg of prednisolone cumulatively, administered in multiple courses for the treatment of exacerbations of COPD.


Conductance through two dots coupled in series: application of the exact solution.
Using the exact Bethe-ansatz solution, we study the conductance through symmetric and asymmetric double-dot (DD) systems in the Kondo regime, where two quantum dots are coupled in series. The application of the Ward-Takahashi identity enables us to compute the conductance at low temperatures. For strong inter- and intra-dot Coulomb repulsions with weak inter-dot tunneling, we discuss how the Kondo effect evolves for an asymmetric DD system as well as a symmetric DD system in a magnetic field. In particular, we clarify that the conductance is decreased (increased) in the asymmetric DD (symmetric DD in a field), reflecting that the Kondo effect due to inter-dot charge fluctuations is suppressed (enhanced).


Choosing a health plan: what information will consumers use?
Employers and policymakers are looking for ways to encourage competition among health plans, thus lowering costs and improving quality. Employers hope to foster competition among health plans by creating standardized measures of quality that supplement the traditional benefits and cost information employees use to compare plans and make choices. This DataWatch examines employees' interest in standardized measures of plan performance. Results from a survey of Massachusetts state employees show that cost and benefit information receive high rankings, but certain plan performance information does not.


Circadian clock control of hepatic lipid metabolism: role of small heterodimer partner (Shp).
Hepatic steatosis, the accumulation of triglyceride droplets in the hepatocytes, is a common hepatic pathology seen in subjects with obesity/metabolic syndrome and those with excessive alcohol use. The pathogenesis underlying hepatic steatosis is complex. Recent studies have shown the specific role played by the molecular clock mechanism in the control of lipid metabolism and that the disruption of these tissue clocks may lead to the disturbances in lipid homeostasis. This review reports a novel role of small heterodimer partner in maintaining triglyceride and lipoprotein homeostasis through neuronal PAS domain protein 2.


Artesunate attenuates glioma proliferation, migration and invasion by affecting cellular mechanical properties.
Glioma is one of the most common malignant brain tumors. Current chemotherapy is far from providing satisfactory clinical outcomes for patients with glioma. More efficient drugs are urgently needed. Artesunate (ART) is clinically used as an anti-malarial agent and exhibits potent antiproliferative activity as a traditional Chinese medicine. In addition, ART has been shown to exert a profound cytotoxic effect on various tumor cell lines, presenting a novel candidate for cancer chemotherapy. However, its anticancer effect on glioma by altering cell biomechanical properties remains unclear. The present study aimed to identify the anticancer effects of ART on human glioma SHG44 cells by assessing cell proliferation, migration/invasion, the expression of claudin-1 and the biomechanical properties of ART-treated SHG44 cells. The proliferation of the SHG44 cells was assessed by MTT assay. The cell apoptosis was detected by flow cytometry. For cell migration and invasion assays, the Transwell was used. The expression of the gene claudin-1 was detected by polymerase chain reaction. The cell membrane and biomechanical properties, as targets of ART action, were investigated by atomic force microscopy (AFM). ART significantly inhibited the proliferation of SHG44 cells in a dose- and time-dependent manner. After treatment with 30 mg/l ART, the level of cell apoptosis was significantly increased (from 6.88±0.062 to 23.7±4.16%). Furthermore, the cell migration and invasion abilities of the SHG44 cells were markedly inhibited after treatment with 30 mg/l ART. Compared with the control group (0 mg/l ART), the SHG44 cells treated with 30 mg/l ART exhibited upregulated expression of claudin-1, increased adhesive force (from 2,400±300 to 3,600±500 pN), increased high connection among SHG44 cells, increased cytomembrane roughness (from 0.118±0.011 to 0.269±0.015 µm) and reduced elasticity (from 23±8 to 3.5±1.1 MPa). The present study demonstrated that ART could alter the biomechanical properties of the glioma cells to inhibit cell proliferation, migration and invasion.


Captopril. An update of its pharmacodynamic and pharmacokinetic properties, and therapeutic use in hypertension and congestive heart failure.
Captopril is an orally active inhibitor of angiotensin-converting enzyme (ACE) and has been widely studied in the treatment of patients with mild to moderate essential hypertension, severe hypertension not responsive to conventional diuretic/beta-adrenoceptor blocker/vasodilator regimens, and patients with chronic congestive heart failure refractory to treatment with a diuretic and digitalis. In patients with mild or moderate essential hypertension, titrated low doses of captopril used alone or in conjunction with a diuretic are similar in efficacy to usual doses of hydrochlorothiazide, chlorthalidone, or beta-adrenoceptor blocking drugs, as well as to the other ACE inhibitors. In addition, captopril improved well-being to a greater extent than methyldopa or propranolol in a study designed specifically to determine the effect of treatment on the quality of life of patients with mild or moderate essential hypertension. The earlier demonstrated efficacy of captopril, used with a diuretic and often also with a beta-adrenoceptor blocking drug, in the treatment of severe hypertension refractory to conventional 'triple therapy' has been confirmed in more recent trials which illustrate the generally marked antihypertensive effect of captopril-containing regimens in such patients. Results of initial trials in patients with scleroderma are promising, with control of hypertension and stabilization of renal function in these patients when treated at an early stage of the disease. Several comparative and long term trials of captopril in patients with chronic congestive heart failure refractory to treatment with a diuretic/digitalis regimen clearly demonstrate that initial haemodynamic improvement is maintained and correlates with clinical benefit. A tendency for overall clinical response to captopril to be better than the response to prazosin, hydralazine, nisoldipine or enalapril has been reported. Results of a multicentre comparison with digoxin and placebo indicate that captopril is a suitable alternative to digoxin in patients with mild to moderate heart failure who are receiving maintenance diuretic therapy. The tolerability of captopril has now been studied in many thousands of patients involved in formalized trials and the early impression of poor tolerability can no longer be justified. The use of generally lower dosages of captopril in patients with normal or slightly impaired renal function has resulted in a generally low incidence of rash (0.5 to 4%), dysgeusia (0.1 to 3%), proteinuria (0.5%), neutropenia (0.3% during first 3 months) and symptomatic hypotension (0.1 to 3%). Cough is an infrequent but troublesome effect resulting from ACE inhibition.(ABSTRACT TRUNCATED AT 400 WORDS)


Capacitance fluctuations causing channel noise reduction in stochastic Hodgkin-Huxley systems.
Voltage-dependent ion channels determine the electric properties of axonal cell membranes. They not only allow the passage of ions through the cell membrane, but also contribute to an additional charging of the cell membrane resulting in the so-called capacitance loading. The switching of the channel gates between an open and a closed configuration is intrinsically related to the movement of gating charge within the cell membrane. At the beginning of an action potential, the transient gating current is opposite to the direction of the current of sodium ions through the membrane. Therefore, the excitability is expected to become reduced due to the influence of a gating current. Our stochastic Hodgkin-Huxley-like modeling takes into account both the channel noise-i.e. the fluctuations of the number of open ion channels-and the capacitance fluctuations that result from the dynamics of the gating charge. We investigate the spiking dynamics of membrane patches of a variable size and analyze the statistics of the spontaneous spiking. As a main result, we find that the gating currents yield a drastic reduction of the spontaneous spiking rate for sufficiently large ion channel clusters. Consequently, this demonstrates a prominent mechanism for channel noise reduction.


An investigation of the neurotrophic factor genes GDNF, NGF, and NT3 in susceptibility to ADHD.
Attention deficit hyperactivity disorder (ADHD) is a common, highly heritable, neurodevelopmental disorder with onset in early childhood. Genes involved in neuronal development and growth are, thus, important etiological candidates and neurotrophic factors have been hypothesized to play a role in the pathogenesis of ADHD. Glial derived neurotrophic factor (GDNF), nerve growth factor (NGF (beta subunit)), and neurotrophic factor 3 (NT3) are members of the neurotrophin family and are involved in the survival, differentiation, and maintenance of neuronal cells. We have examined 10 coding and intronic single nucleotide polymorphisms (SNPs) across GDNF, NGF, and NT3 in a family-based association sample of 120 DSM-IV ADHD probands and their biological parents, as well as a case-control analysis with 120 sex-matched controls. Borderline significant overtransmission of the C allele of a non-synonymous C/T SNP (rs6330) in NGF which codes an alanine/valine change was found in the family-based sample (Chi-square = 3.69, odds ratio (OR) = 1.65, P = 0.05). Although this SNP is located in the 5' pro-NGF sequence and not the mature NGF protein, it may affect intracellular processing and secretion of NGF.


The prognostic value of the number of negative lymph nodes in esophageal cancer patients after transthoracic resection.
Although the number of positive lymph nodes (LN), the total number of resected LN, and the proportion of positive LN have been reported to be associated with survival in patients with esophageal cancer, little is known about the prognostic impact of the number of negative LN. A retrospective review of 754 patients receiving transthoracic esophagectomy for cancer between January 1995 and September 2011 was performed. The prognostic impact of the number of negative LN was analyzed. Log rank testing was used to compare survival curves, and Cox regression analysis was performed to identify significant prognostic factors. A total of 707 patients were included. The mean follow-up time was 28.4±30.9 months. The 5-year overall survival for the entire cohort was 30.1%. Patients with a high number of negative LN (≥19) had better overall survival than patients with a low number of negative LN (5-year survival rate 33.4% versus 26.4%, p=0.005). Stratified analysis showed that the impact of the number of negative LN was significant in patients with T3/4 (p=0.027) and node-positive (p=0.002) esophageal cancers but not in patients with less advanced tumors. Multivariate Cox regression analysis demonstrated that the number of negative LN (in addition to age, sex, T stage, N stage, tumor length, and surgical approach) was an independent prognostic factor. A higher number of negative LN is associated with better overall survival of esophageal cancer patients after resection. The correlation of a high number of negative LN (≥19) with survival was more prominent in patients with advanced (T3/4 stage, node-positive) tumors.


Fixation of Clostridium difficile toxin A and cholera toxin to intestinal brush border membranes from axenic and conventional mice.
We have tested the in vitro binding of Clostridium difficile toxin A (enterotoxin) and cholera toxin to intestinal brush border membranes prepared from either conventional or axenic mice. Membranes from axenic mice were shown to be saturated at a lower toxin A concentration (at least 2.5 times lower). Because there were no significant differences between membranes from axenic and conventional mice in binding at low toxin A concentrations, the presence of the normal microflora seems to increase the number but not the affinity of brush border membrane receptors on the enterocyte surface. Corroborating the in vitro results, we observed that conventional mice were more sensitive to the pathological effects of toxin A given intragastrically than were axenic mice. In contrast, there was no difference in the binding characteristics of cholera toxin between membranes from conventional and axenic mice. We conclude that the presence of the mouse intestinal bacteria increases the number of C. difficile toxin A intestinal receptors but does not influence cholera toxin receptors.


Potentiation of neurovirulence of canine distemper virus in guinea pigs by sensitization with neural antigen.
Establishment of animal model of virus-induced encephalomyelitis was attempted in strain 13 guinea pigs sensitized with the homologous spinal cord antigen. Intracerebral inoculation of canine distemper virus alone or sensitization with the neural antigen alone did not induce significant clinical signs. Mild histological lesions were found in both the meninges and parenchyma of the central nervous system (CNS) of virus-infected animals, and in the meninges of the sensitized ones. In contrast, combination of virus infection and sensitization resulted in development of neurological signs of CNS disease as well as of marked histological lesions involving both the meninges and parenchyma. The potentiated CNS disease was successfully transferred by the lymph node cells of the sensitized animals into virus-infected ones. These results suggested that the virus-induced lesions in the CNS were potentiated by the lymphocytes sensitized with neural antigen. Depending on the time schedule of the sensitization and virus infection, different courses of CNS diseases including acute, subacute, and/or recurrent ones were induced, indicating the usefulness of this animal model for immunological and virological analysis of virus-induced CNS diseases.


Wear-simulation analysis of rotating-platform mobile-bearing knees.
The wear and wear debris from rotating-platform mobile-bearing knees and fixed-bearing knees were compared in knee joint-simulator studies. The wear rate of the fixed-bearing knees was found to increase as the kinematics were increased because of an increase in internal-external rotation and an increase in anterorposterior (AP) translation. The wear rate of the rotating-platform mobile-bearing knees was found to be significantly lower than that of the fixed-bearing knees. The rotating-platform mobile-bearing knee was able to decouple the complex kinematics to pure rotation at the inferior tibial articulating surface and linear flexion-extension and AP sliding at the superior femoral articulating interface, substantially reducing cross-shear and wear. No difference was found in the wear debris between the rotating-platform and fixed-bearing knees. This resulted in a substantially reduced functional biological activity or osteolytic potential for the rotating-platform mobile-bearing knees due to the lower wear rates.


Processing of Plasmodium falciparum Merozoite Surface Protein MSP1 Activates a Spectrin-Binding Function Enabling Parasite Egress from RBCs.
The malaria parasite Plasmodium falciparum replicates within erythrocytes, producing progeny merozoites that are released from infected cells via a poorly understood process called egress. The most abundant merozoite surface protein, MSP1, is synthesized as a large precursor that undergoes proteolytic maturation by the parasite protease SUB1 just prior to egress. The function of MSP1 and its processing are unknown. Here we show that SUB1-mediated processing of MSP1 is important for parasite viability. Processing modifies the secondary structure of MSP1 and activates its capacity to bind spectrin, a molecular scaffold protein that is the major component of the host erythrocyte cytoskeleton. Parasites expressing an inefficiently processed MSP1 mutant show delayed egress, and merozoites lacking surface-bound MSP1 display a severe egress defect. Our results indicate that interactions between SUB1-processed merozoite surface MSP1 and the spectrin network of the erythrocyte cytoskeleton facilitate host erythrocyte rupture to enable parasite egress.


[Paratesticular rhabdomyosarcoma. A case report].
The rhabdomyosarcomas are rare tumors of mesenchymal tIssue. Whose most frequent localisations are génito-urinary, and occur in the child and young adult. It's an intrascrotal tumour, localised in the tunica vaginalis, epididymis or spermatic cord. The Authors report a case of paratesticular rhabdomyosarcoma in a 15 Year old child, who presented a right painless scrotal mass since two Months. The research of the tumoral markers is negative. Scrotal ultrasound shows a tissular mass in spermatic cord. Abdominal ultrasound and CT show retroperitoneal lymph node mass. Chest X-ray is normal. The intervention consists of a right castration. The histological examination of the spiceman confirms the diagnosis of the paratesticular rhabdomyosarcoma of the spermatic cord. The patient is treated by chemotherapy without improvement. Through this observation, the Authors underline the interest of the imagery and the aspects of the paratesticular tumours in ultrasound and CT.


Nonlethal gamma radiation displays similar cytoplasmic rearrangements as 12(S)-HETE.
Timar et al. (Int. J. Canc., 52:594-603, 1992) and Onoda et al. (Rad. Res., 130:281-288, 1992) implicated 12(S)-HETE and nonlethal gamma radiation respectively, in enhanced expression of the alpha IIb beta 3 integrin receptor in B16a tumor cells. 12(S)-HETE has been reported to induce rearrangement of cytoplasmic elements and movement of cytoplasm (Haddad and Taylor, Proc. Amer. Cancer Res., 34:51, 1993); Timar et al., Cell Motil. Cytoskel., 26:49-65, 1993). Here, we report similar results in B16a tumor cells by post 90 cGy gamma radiation. Control cells were circular without cytoplasmic processes with the majority of their organelles and elements in perinuclear regions both surrounded by a ring of actin microfilaments. The cell's extensive leading edge was organelle free with the exception of occasional endoplasmic reticular cisternae parallel to one or two microtubules, and short bundles of actin microfilaments perpendicular to the plasma membrane. As early as 5 sec, but better defined by 60-90 sec, one end of elongated microfilaments bundles became associated with the plasma membrane while the other end of the bundles became associated with perinuclear mass protrusions. By 5 min, prominent by 15 min, protrusions, now separated from the leading edge by mixture of bundle and ring microfilaments, reached the plasma membrane to form cellular processes. By 30 min, some cytoplasmic masses showed signs of retraction along with the disappearance of their associated mixed microfilaments. As well as numerous vesicles of unknown origin, primarily associated with microtubules perpendicular to the plasma membrane, mitochondria and endoplasmic reticular cisternae were found in the cytoplasmic masses.


Contralateral Effect following Intravitreal Brolucizumab Injection in Diabetic Macular Edema.
The authors describe a novel case of a 48-year-old male with bilateral diabetic macular edema (DME) who underwent intravitreal injection (IVI) of brolucizumab in the left eye. At four weeks, the patient demonstrated a bilateral response by way of improvement in the best-corrected visual acuity (BCVA) and reduction in the central macular thickness (CMT) in both eyes. Further studies on the ocular and systemic assays of the brolucizumab molecule are warranted to evaluate its systemic escape and to better understand the pharmacokinetics behind the bilateral effect.


Thyroid nodule with Hashimoto thyroiditis in childhood - a challenging experience.
Fine-needle aspiration biopsy (FNAB) of the thyroid, although not perfect, is considered currently the best preoperative method in establishing malignancy in a thyroid nodule. We report the case of an 8-year-old girl with a thyroid mass in the right thyroid lobe. Endocrine investigations revealed subclinical hypothyroidism and high titer of anti-thyroid peroxidase antibodies. Fine Needle Aspiration Biopsy (FNAB) of the right side of the thyroid gland, where on thyroid ultrasonography (US) a poorly defined iso-/hypoechoic nodule with irregular margins was detected, revealed a background of lymphocytes and plasma cells mixed with follicular cells with reactive changes. After four months of treatment with L-thyroxine, ultrasonographic follow up showed an increase in nodule size. FNAB was performed again but showed non-diagnostic cytology. Right lobectomy was performed and the pathology report shows papillary carcinoma, follicular variant predominantly with components of insular growth. We discuss the false-negative FNAB interpretation because of the concomitant autoimmune thyroiditis. Management strategy for this case and the prognosis is also discussed.


Environmental filtering and phylogenetic clustering correlate with the distribution patterns of cryptic protist species.
The community composition of any group of organisms should theoretically be determined by a combination of assembly processes including resource partitioning, competition, environmental filtering, and phylogenetic legacy. Environmental DNA studies have revealed a huge diversity of protists in all environments, raising questions about the ecological significance of such diversity and the degree to which they obey to the same rules as macroscopic organisms. The fast-growing cultivable protist species on which hypotheses are usually experimentally tested represent only a minority of the protist diversity. Addressing these questions for the lesser known majority can only be inferred through observational studies. We conducted an environmental DNA survey of the genus Nebela, a group of closely related testate (shelled) amoeba species, in different habitats within Sphagnum-dominated peatlands. Identification based on the mitochondrial cytochrome c oxidase 1 gene, allowed species-level resolution as well as phylogenetic reconstruction. Community composition varied strongly across habitats and associated environmental gradients. Species showed little overlap in their realized niche, suggesting resource partitioning, and a strong influence of environmental filtering driving community composition. Furthermore, phylogenetic clustering was observed in the most nitrogen-poor samples, supporting phylogenetic inheritance of adaptations in the group of N. guttata. This study showed that the studied free-living unicellular eukaryotes follow to community assembly rules similar to those known to determine plant and animal communities; the same may be true for much of the huge functional and taxonomic diversity of protists.


Influences of cognitive control on numerical cognition--adaptation by binding for implicit learning.
Recently, an associative learning account of cognitive control has been suggested (Verguts & Notebaert, 2009). In this so-called adaptation by binding theory, Hebbian learning of stimulus-stimulus and stimulus-response associations is assumed to drive the adaptation of human behavior. In this study, we evaluated the validity of the adaptation-by-binding account for the case of implicit learning of regularities within a stimulus set (i.e., the frequency of specific unit digit combinations in a two-digit number magnitude comparison task) and their association with a particular response. Our data indicated that participants indeed learned these regularities and adapted their behavior accordingly. In particular, influences of cognitive control were even able to override the numerical distance effect--one of the most robust effects in numerical cognition research. Thus, the general cognitive processes involved in two-digit number magnitude comparison seem much more complex than previously assumed. Multi-digit number magnitude comparison may not be automatic and inflexible but influenced by processes of cognitive control being highly adaptive to stimulus set properties and task demands on multiple levels.


The California Prenatal Screening Program: "options and choices" not "coercion and eugenics".
The California Prenatal Screening Program is designed to make prenatal screening available to the state's large and diverse population. The Program provides information to women which will allow them to make informed choices regarding prenatal screening and prenatal diagnosis. Since the Program's inception in 1986, women in California have had the option to participate in prenatal screening or to decline prenatal screening. The California Program offers prenatal diagnostic services to women whose screening tests indicate an increased risk for birth defects, including Down syndrome. Women can decline any or all of these follow-up services. Genetic counseling, diagnostic services, and the presentation of diagnostic results are performed by medical professionals (not State staff) who follow established guidelines for nondirective counseling. Program data clearly demonstrate that women in California have a wide range of options and make a wide range of choices regarding prenatal screening and prenatal diagnosis. California's comprehensive Prenatal Screening Program promotes optimal care for all women within all options and choices. The important and necessary communication among organizations and stakeholders involved in prenatal screening and diagnosis, and in related care for pregnant women and for people with Down syndrome, is not served by misrepresentation and inflammatory rhetoric.


ONTOGENETIC VARIATION IN PATTERNS OF DEVELOPMENTAL AND FUNCTIONAL INTEGRATION IN SKULLS OF SIGMODON FULVIVENTER.
Patterns of variation and covariation within populations can influence how characters respond to natural selection and random genetic drift and so constrain the ability of natural selection to modify the phenotype. We examined several potential developmental and functional explanations of character covariation throughout ontogeny using known-age samples of the cotton rat (Sigmodon fulviventer) to identify the causes of covariation and to assess the variability of patterns of covariation throughout postnatal growth. Competing developmental and functional models were fit to samples of orofacial and neurocranial measures by confirmatory factor analysis and evaluated for their ability to reconstruct observed variance-covariance matrices. Samples of successive ages were simultaneously fit to a common model to test the hypothesis that the patterns of developmental and functional integration were invariant between ages. Orofacial characters derived from the same branchial-arch primordium covary early in ontogeny. Subsequently, there is a repatterning of integration that may reflect a transition from developmental to functional sources of integration. Neurocranial characters exhibit even more variation in patterns of covariation: initially, characters appear to comprise a single integrated unit; before puberty, they appear to respond to localized bone growth; after puberty, they form separate calvarial and basicranial components. This ontogenetic variation in patterns of covariation suggests that developmental constraints are transient and flexible and that the consequences of selection may depend upon the age at which it acts.


Prevalence and determinants of HIV in tuberculosis patients in Wuxi City, Jiangsu province, China: a cross-sectional study.
At least one-third of the 34 million people living with human immunodeficiency virus (HIV) worldwide are infected with latent tuberculosis (TB). The aim of this study was to determine the rate of HIV infection in TB patients and its determinants in Wuxi City, China. TB patients attending health institutions (12 selected sites) for TB diagnosis and treatment were enrolled in this study. TB diagnosis, treatment and HIV testing were done according to the national guidelines. Blood samples were collected for anonymous HIV testing. Among the TB patients, the HIV prevalence was 13.66% (1493/10,926). Multivariate analysis showed that gender, age, education, marital status, per capita monthly income, patient residence, family size, distance from a health institution, knowledge of HIV-TB co-infection, and knowledge of HIV may be risk factors for HIV-TB co-infection (all: odds ratio > 1, p < 0.05). The prevalence of TB in those with HIV was higher among the study participants. Improving public awareness of HIV-TB co-infection, regularly screening and improving follow-up can reduce the occurrence of HIV-TB co-infection.


Isolation and characterization of an Escherichia coli DnaK mutant with impaired ATPase activity.
A temperature-sensitive mutant of DnaK, the principal Escherichia coli member of the 70 kDa heat shock protein family, has been isolated. The mutation, dnaK25, lies in the putative ATP binding pocket of DnaK. It consists of a C to T transition that changes the highly conserved proline 143 to serine. Mutant strains do not support the propagation of bacteriophage lambda or of plasmids that require DnaA for replication. They are also defective in the utilization of mannose and sorbitol. ATPase activity of the mutant protein is reduced 20-fold relative to wild-type, while autophosphorylation is unaffected. DnaK25 has a fourfold faster rate of nucleotide exchange than wild-type DnaK; nucleotide exchange by both proteins is markedly increased by GrpE. The DnaK25 ATPase is still stimulated by DnaJ and GrpE and by peptide substrates. However, the affinity of most peptides tested for stimulating the DnaK25 ATPase is reduced significantly. These results indicate that a mutation in the N-terminal nucleotide binding domain can alter substrate interactions with the C-terminal substrate binding site. Nucleotide exchange by both wild-type DnaK and DnaK25 proceeds at a much faster rate than ATP hydrolysis, and therefore cannot be the rate limiting step of ATP hydrolysis under the conditions used in these experiments. Consistent with this, peptides, which stimulate ATP hydrolysis, have no effect on nucleotide exchange. Peptides thus appear to stimulate the ATPase by acting at another step, such as increasing the rate of phosphate bond cleavage.


Endosomal localization and function of sorting nexin 1.
There are 17 human members of the sorting nexin (SNX) family of proteins that contain Phox (PX) domains. Yeast orthologs function in vesicular trafficking and mammalian proteins have been implicated in endocytic trafficking of cell surface receptors. The first member of this family, SNX1, was identified via interaction with the epidermal growth factor receptor. The present studies indicate that SNX1 and SNX2 are colocalized to tubulovesicular endosomal membranes and this localization depends on PI 3-kinase activity. Point mutations in the PX domain that abolish recognition of phosphorylated phosphatidylinositol (PtdIns) in vitro abolish vesicle localization in vivo indicating that lipid binding by the PX domain is necessary for localization to vesicle membranes. Deletion of a predicted coiled-coil region in the COOH terminus of SNX1 also abolished vesicle localization, indicating that this helical domain, too, is necessary for SNX1 localization. Thus, both PX domain recognition of PtdIns and COOH terminal helical domains are necessary for localization of SNX1 with neither alone being sufficient. Regulated overexpression of the NH(2) terminus of SNX1 containing the PX domain decreased the rate of ligand-induced epidermal growth factor receptor degradation, an effect consistent with inhibition of endogenous SNX1 function in the endosome compartment. SNX1 thus functions in regulating trafficking in the endosome compartment via PX domain recognition of phosphorylated PtdIns and via interaction with other protein components.


Leveraging Technology for Productivity and Best Outcomes.
The case described herein, illustrates the comfortable interplay that can be achieved between traditional techniques and new technologies. At the patient's first visit in the dental office, traditional tools can be used to gather records, and an old-fashioned dialogue was employed to determine the patient's goals for treatment. Creation of a digital treatment simulation by TransLab gives the dentist, dental laboratory team, and the patient a tangible picture to review and discuss. This collaboration helps to ensure that all parties have the same expectations, before proceeding any further with the actual treatment procedures.


Influence of ADP-ribosyltransferase inhibitors on the intracellular NAD and ATP levels in Ehrlich ascites tumor cells: implication for the altered NAD + ATP-dependent cellular sensitivity to the cytotoxic agents.
Exposure of Ehrlich ascites tumor cells to 3-aminobenzamide for 60 min resulted in a dose-dependent increase of cellular NAD and ATP levels at a concentration range of 0.3-5 mM. In the cells exposed to 5-methylnicotinamide there was a decrease of both nucleotide levels. As a possible cause for these changes we found a marked inhibition of microsomal NAD glycohydrolase activity by 3-aminobenzamide and a moderate stimulation of this enzyme by 5-methylnicotinamide. Furthermore, 3-aminobenzamide significantly enhanced the cellular uptake of nicotinamide and NAD synthesis, probably by the stimulation of nuclear ATP-NMN adenylyltransferase activity. We show also that the cells containing elevated NAD and ATP levels by the exposure to 3-aminobenzamide became resistant to the 5-azacytidine cytotoxicity.


Modulation of toll-like receptor 4. Insights from x-ray crystallography and molecular modeling.
Toll-like receptors (TLRs) are a family of proteins with a key role in the innate immune system. They are specialized in the recognition of molecular patterns present in microbial components, through mechanisms not yet unraveled at atomic level. Improvement in the understanding of the molecular mechanisms that drive TLR signaling is of paramount importance to grasp key aspects of immunity, potentially leading to the design of new molecules able to modulate their functions. Toll-like receptor 4 (TLR4), along with its accessory protein myeloid differentiation factor 2 (MD-2), builds a heterodimeric complex that specifically recognizes lipopolysaccharides (LPS), which are present on the cell wall of gramnegative bacteria, activating the immune response. Some TLR4 modulators are undergoing preclinical and clinical evaluation for the treatment of sepsis, inflammatory diseases, cancer, and rheumatoid arthritis. Reported X-ray crystal structures together with molecular modeling studies, not reviewed before in the literature, have recently contributed to the elucidation of key interactions at atomic level of the binding between the TLR4/MD-2 system and different TLR4/MD-2 ligands. The purpose of this review is to summarize these reported studies which may account for the SAR rationalization of natural/ synthetic agonist/antagonist TLR4 binders and may also guide further design of novel TLR4 modulators.


Astrocyte Senescence and Alzheimer's Disease: A Review.
Astrocytes are the largest group of glial cells in the brain and participate in several essential functions of the central nervous system (CNS). Disruption of their normal physiological function can lead to metabolism disequilibrium and the pathology of CNS. As an important mechanism of aging, cellular senescence has been considered as a primary inducing factor of age-associated neurodegenerative disorders. Senescent astrocytes showed decreased normal physiological function and increased secretion of senescence-associated secretory phenotype (SASP) factors, which contribute to Aβ accumulation, tau hyperphosphorylation, and the deposition of neurofibrillary tangles (NFTs) in Alzheimer's disease (AD). Astrocyte senescence also leads to a number of detrimental effects, including induced glutamate excitotoxicity, impaired synaptic plasticity, neural stem cell loss, and blood-brain barrier (BBB) dysfunction. In this review article, we have summarized the growing findings regarding astrocyte senescence and its putative role in the pathologic progress of AD. Additionally, we also focus on the significance of targeting astrocyte senescence as a novel and feasible therapeutic approach for AD.


Benefits and harms of breast cancer screening with mammography in women aged 40-49 years: A systematic review.
Early detection of breast cancer through screening can lower breast cancer mortality rates and reduce the burden of this disease in the population. In most western countries, mammography screening starting from age 50 is recommended. However, there is debate about whether breast cancer screening should be extended to younger women. This systematic review provides an overview of the evidence from RCTs on the benefits and harms of breast cancer screening with mammography in women aged 40-49 years. The quality of the evidence for each outcome was appraised using the GRADE approach. Four articles reporting on two different trials-the Age trial and the Canadian National Breast Screening Study-I (CNBSS-I)-were included. The results showed no significant effect on breast cancer mortality (Age trial: RR 0.93 (95% CI 0.80-1.09); CNBSS-I: HR 1.10 (95% CI 0.86-1.40)) nor on all-cause mortality (RR 0.98, 95% CI 0.93-1.03) in women aged 40-49 years offered screening. Among regularly attending women, the cumulative risk of experiencing a false-positive recall was 20.5%. Over-diagnosis of invasive breast cancer at 5 years post-cessation of screening for women aged 40-49 years was estimated to be 32% and at 20 years post-cessation of screening to be 48%. Including ductal carcinoma in situ, these numbers were 41% and 55%. Based on the current evidence from randomised trials, extending mammography screening to younger age groups cannot be recommended. However, there were limitations including relatively low sensitivity of screening and screening attendance, insufficient power, and contamination, which may explain the nonsignificant results.


A gradient inverse planning algorithm with dose-volume constraints.
An inverse planning algorithm for determining the intensity-modulated beams that will most closely generate a desired dose distribution is presented. The algorithm is three-dimensional and does not explicitly depend on beam energies and modalities. It allows a single prescription dose or a window of acceptable doses to be specified for the target, with additional constraints to account for under- or over-dosing. For the protection of organs at risk, it provides maximum-dose and dose-volume constraints. The latter apply to the entire volume of the organ exposed to the corresponding dose levels. Several levels of each type of constraint, with varying penalty weights, may be specified for each organ. The objective function that serves as the measure of the goodness of the solution is of the least-squares type and is minimized using conjugate gradient methods. Typical clinical cases involving 40,000 points and 4000 rays to be determined require about 10 min of CPU time on a DEC AlphaStation. Results are presented for two clinical sites, prostate and lung. The optimization algorithm yielded plans that featured higher target dose homogeneity, compared with the human planner's plan, while selectively sparing more of the normal organs at the desired dose regions.


Organotypic modeling of human keratinocyte response to peroxisome proliferators.
Peroxisome proliferators (PPs) are a diverse chemical group including hypolipidemic drugs and some fatty acids. Their stimulation of PP-activated receptors (PPARs) and subsequent control of gene expression regulates metabolism and differentiation in many cells. PPs have multiple opportunities to target human epidermal keratinocytes because of delivery through dietary, clinical, and/or topical exposure routes. PPAR knockout mice and PP treatment of mouse skin or human keratinocytes in monolayer culture have established some effects for PPs in cutaneous differentiation. However, incomplete epidermal maturation characteristic of monolayer keratinocytes and rodent-specific effects may limit our full understanding of human keratinocyte responses to PPs. To address these issues, we investigated PP influence on primary human keratinocytes in organotypic cultures that recapitulate biochemical markers of epidermis. We found that the PPARα agonists clofibrate, docasohexaenoic acid, and WY-14,643 produced mild to moderate keratinocyte hyperplasia, increased stratification (particularly of granular and cornified layers), and enhanced levels of the differentiation markers filaggrin, ABCA12, and phosphorylated HSP27. Several PP effects generated in the organotypic system, however, were distinct from those previously reported for rodent skin and human keratinocyte monolayer cultures, suggesting that the species and growth context of target cells can impact exposure outcomes. Given the utility of organotypic cultures for modeling the epidermis, studies in this system may bridge the gap between the rodent assays and clinical studies of human epidermal responses to PPs.


Searching Missing Proteins Based on the Optimization of Membrane Protein Enrichment and Digestion Process.
A membrane protein enrichment method composed of ultracentrifugation and detergent-based extraction was first developed based on MCF7 cell line. Then, in-solution digestion with detergents and eFASP (enhanced filter-aided sample preparation) with detergents were compared with the time-consuming in-gel digestion method. Among the in-solution digestion strategies, the eFASP combined with RapiGest identified 1125 membrane proteins. Similarly, the eFASP combined with sodium deoxycholate identified 1069 membrane proteins; however, the in-gel digestion characterized 1091 membrane proteins. Totally, with the five digestion methods, 1390 membrane proteins were identified with ≥1 unique peptides, among which 1345 membrane proteins contain unique peptides ≥2. This is the biggest membrane protein data set for MCF7 cell line and even breast cancer tissue samples. Interestingly, we identified 13 unique peptides belonging to 8 missing proteins (MPs). Finally, eight unique peptides were validated by synthesized peptides. Two proteins were confirmed as MPs, and another two proteins were candidate detections.


Automated quantification of glenoid bone loss in CT scans for shoulder dislocation surgery planning.
Estimation of glenoid bone loss in CT scans following shoulder dislocation is required to determine the type of surgery needed to restore shoulder stability. This paper presents a novel automatic method for the computation of glenoid bone loss in CT scans. The model-based method is a pipeline that consists of four steps: (1) computation of an oblique plane in the CT scan that best matches the glenoid face orientation; (2) selection of the glenoid oblique CT slice; (3) computation of the circle that best fits the posteroinferior glenoid contour; (4) quantification of the glenoid bone loss. The best-fit circle is computed with newly defined Glenoid Clock Circle Constraints. The pipeline and each of its steps were evaluated on 51 shoulder CT scans (44 patients). Ground truth oblique slice, best-fit circle, and glenoid bone loss measurements were obtained manually from three clinicians. The full pipeline yielded a mean absolute error (%) for the bone loss deficiency of 2.3 ± 2.9 mm (4.67 ± 3.32%). The mean oblique CT slice selection difference was 1.42 ± 1.32 slices, above the observer variability of 1.74 ± 1.82 slices. The glenoid bone loss deficiency measure (%) on the ground truth oblique glenoid CT slice has a mean average error of 0.54 ± 1.03 mm (4.76 ± 3.00%), close to the observer variability of 0.93 ± 1.40 mm (2.98 ± 4.97%). Our pipeline is the first fully automatic method for the quantitative analysis of glenoid bone loss in CT scans. The computed glenoid bone loss report may assist orthopedists in selecting and planning surgical shoulder dislocation procedures.


Particle characteristics in the reactor and pelletizing areas of carbon black production.
Physical and chemical characteristics of airborne particles (ultrafine, PM1, PM2.5, and PM10) in reactor and pelletizing areas during carbon black production were measured to assess process related sources of particles in work areas. Results from bagging areas within the same three facilities have been previously published. Particle number and mass concentration measurements were conducted in these work areas and at ambient comparison sites at each of the three carbon black plants. No elevated ultrafine particle number concentrations (UFP, <100 nm) with respect to ambient were determined in the work areas of Plant 1, intermittently elevated concentrations at Plant 2, and permanently elevated concentrations at Plant 3. The intermittently elevated UFP concentrations in the pelletizer and reactor areas of Plant 2 could be related to nearby traffic emissions. The ultrafine particle number concentrations at Plant 2 are comparable to those determined at urban traffic sites. Both work areas of Plant 3 showed elevated UFP concentrations in the pelletizer reactor and areas. In the case of the reactor, which was the only enclosed reactor area investigated among the three facilities, the source of the elevated UFP number concentration was most likely attributable to grease and oil fumes from maintenance activities, a conclusion supported by carbon fractionation analysis. The elevated UFP number concentrations in the pelletizing area in this same plant are related to leaks in the production line, which allowed particulate matter to escape to the surrounding areas. Absolute PM10 mass concentrations were all within normal ambient concentrations except for the pelletizing area in Plant 3, which showed continuous levels above ambient. One additional source contributing to peak level PM10 mass concentrations at Plant 2 was due to wind dispersion from a carbon black spill incident the day prior to measurements. It is concluded from these measurements that no carbon black is released in the reactor and pelletizing areas (as UFP or PM10) from the closed production lines under normal operating conditions.


Anticancer Dose Adjustment for Patients with Renal and Hepatic Dysfunction: From Scientific Evidence to Clinical Application.
Most anticancer agents exhibit a narrow therapeutic index, i.e., a small change in plasma concentrations can lead to a less efficacious treatment or an unacceptable degree of toxicity. This study aimed at providing health professionals with a feasible and time-saving tool to adapt the dose of anticancer agents for patients with renal or hepatic dysfunction. A guideline for anticancer agents was developed based on a literature search. An algorithm was generated to enhance the efficiency of the dose adaptation process. Finally, the dosing guideline was converted into an easy-to-use ExcelTM tool. The concept was applied to a total of 105 adult patients at the Centre for Integrated Oncology, Bonn, Germany. In total, 392 recommendations for dose adaptation were made and 320 (81.6%) recommendations were responded to by the oncologists. 98.4% of the recommendations were accepted. The algorithm simplifies the decision and screening process for high-risk patients. Moreover, it provides the possibility to quickly decide which laboratory tests are required and whether a dose adjustment for a particular anticancer drug is needed. The ExcelTM tool provides a recommended individual dose for patients with renal or hepatic dysfunction. The effectiveness of this strategy to reduce toxicity should be investigated in further studies before being adopted for routine use.


What Role Does CFTR Play in Development, Differentiation, Regeneration and Cancer?
One of the key features associated with the substantial increase in life expectancy for individuals with CF is an elevated predisposition to cancer, firmly established by recent studies involving large cohorts. With the recent advances in cystic fibrosis transmembrane conductance regulator (CFTR) modulator therapies and the increased long-term survival rate of individuals with cystic fibrosis (CF), this is a novel challenge emerging at the forefront of this disease. However, the mechanisms linking dysfunctional CFTR to carcinogenesis have yet to be unravelled. Clues to this challenging open question emerge from key findings in an increasing number of studies showing that CFTR plays a role in fundamental cellular processes such as foetal development, epithelial differentiation/polarization, and regeneration, as well as in epithelial-mesenchymal transition (EMT). Here, we provide state-of-the-art descriptions on the moonlight roles of CFTR in these processes, highlighting how they can contribute to novel therapeutic strategies. However, such roles are still largely unknown, so we need rapid progress in the elucidation of the underlying mechanisms to find the answers and thus tailor the most appropriate therapeutic approaches.


[Poorly differentiated carcinoma of the thyroid--immunohistochemical evaluation of p53 and p27 protein expression].
Thyroid carcinomas represent a broad spectrum of tumours with different biologic behaviours. The gap between the indolent course of well-differentiated papillary and follicular carcinomas and the very aggressive behaviour of anaplastic carcinomas is filled by variants of thyroid carcinoma with intermediate prognosis, designated in the literature as poorly differentiated (with insular, trabecular and solid patterns) and tall/columnar cell variant of papillary carcinoma. The study has been carried out in 103 patients who had thyroid cancer with various grade of differentiation. The diagnostic significance of p53 and p27 expression in tumor cells has been investigated by immunohistochemical analysis. Well differentiated carcinomas have exhibited the lowest p53 staining frequency, the expression has been higher in poorly differentiated carcinomas and 100% of anaplastic carcinomas have been positive p27 immunohistochemistry has been positive in 80% of investigated carcinomas (in 15% strong positive reaction). In the group of poorly differentiated tumours positive reaction has been observed in 75% cases (in 15% strong positive reaction). Our data suggest that expression of p53 and p27 seems to have limited routine diagnostic significance, but p53 positivity is a good marker of tumour progression.


BET, a novel neuronal transmembrane protein with multiple EGF-like motifs.
Using the signal sequence trap method, we have identified a cDNA clone coding for a type I transmembrane protein, BET, with 10 epidermal growth factor (EGF) motifs in the extracellular domain. In situ hybridization revealed that the bet mRNA is specifically expressed in the mitral/tufted cells in the olfactory bulb, Purkinje cells in the cerebellum, and pyramidal cells in the hippocampus. Using polyclonal antibodies, we have demonstrated that the BET protein is highly glycosylated and is localized in patches in the dendrites and in the somata of neurons. Since the predicted structure of BET shares many similarities with the Notch ligands, this novel protein may play crucial roles in establishing the neuronal networks in the olfactory system, cerebellum, and hippocampus. BET is the first transmembrane protein containing only multiple EGF-like repeats specifically expressed in the brain.


Digital story telling in social justice nursing education.
The purpose of this study was to explore and evaluate how digital stories integrated into public health nursing education can teach social justice concepts essential for nurse leadership. Four digital stories were selected and incorporated into a public health nursing course. Students were asked to reflect on these stories. A retrospective qualitative analysis was completed on the student narrative reflections and analyzed for themes. A total of 108 narrative reflections of public health nursing students were included from 2015 to 2016. Themes were identified based on analysis and include-Encountering Vulnerability, Questioning Systems and Choosing Moral Courage. Digital stories offer an innovative medium to convey the importance of story, advance social justice as an essential practice of nursing, and create opportunities that addresses social justice in nursing and in developing nursing leaders.


Sources and dynamics of fluorescent particles in hospitals.
Fluorescent particles can be markers of bioaerosols and are therefore relevant to nosocomial infections. To date, little research has focused on fluorescent particles in occupied indoor environments, particularly hospitals. In this study, we aimed to determine the spatial and temporal variation of fluorescent particles in two large hospitals in Brisbane, Australia (one for adults and one for children). We used an Ultraviolet Aerodynamic Particle Sizer (UVAPS) to identify fluorescent particle sources, as well as their contribution to total particle concentrations. We found that the average concentrations of both fluorescent and non-fluorescent particles were higher in the adults' hospital (0.06×106 and 1.20×106 particles/m3 , respectively) than in the children's hospital (0.03×106 and 0.33×106 particles/m3 , respectively) (P<.01). However, the proportion of fluorescent particles was higher in the children's hospital. Based on the concentration results and using activity diaries, we were able to identify sources of particle production within the two hospitals. We demonstrated that particles can be easily generated by a variety of everyday activities, which are potential sources of exposure to pathogens. Future studies to further investigate their role in nosocomial infection are warranted.


Dissection of the roles of adenine ring nitrogen (N-1) and exocyclic amino (N-6) moieties in the interaction of 2-5A with RNase L.
To elucidate further the roles played by the adenine bases in the interaction of RNase L (EC 220.127.116.11) with the 2',5'-oligoadenylate 2-5A, p5'A2'(p5'A2')np5' A, a series of sequence-specific 1-deazaadenosine (c1A)-substituted analogues were synthesized and evaluated for their ability to bind to and activate human RNase L in comparison to earlier reported inosine-substituted congeners of 2-5A. Substitution of only the 5'-terminal adenosine of p5'A2'p5'A2 p5 A with c1A afforded an analogue with strongly diminished RNase L binding and activation ability, while replacement of the second or middle adenosine of p5 A2' p5'A2'p5' A had only a modest effect. In distinct contrast to p5'A2'p5'A2'p5'I, the c1A analogue with the third or 2'-terminal adenosine replacement approached parent p5' A2'p5'A2'p5' A in RNase L activation ability. These results permitted a further dissection of the role of various nucleotidic functional groups in the interaction of 2-5A with RNase L: specifically, that the 5'-terminal adenosine purine N-1 moiety is key for binding to RNase L, while the 2'-terminal adenosine N-6 exocyclic amino group is critical for RNase L activation.


Reduced first-phase insulin secretion increases postprandial lipidemia in subjects with impaired glucose tolerance.
It is not clear which phase of insulin secretion is more important to regulate lipoprotein lipase (LPL) activity. After a meal, insulin is released and acts as a major regulator of LPL activity. Postprandial hyperlipidemia is a common comorbidity in subjects with insulin resistance (IR). Therefore this study aimed to evaluate the role of the first-phase insulin secretion (FPIS) on postprandial lipidemia in subjects with IR and impaired glucose tolerance (IGT). This is a cross-sectional, observational and comparative study. We included male and female subjects between 40 and 60 years with a body mass index (BMI) between 23 and 30 kg/m2. Then, patients were divided into three groups. Group 1 consisted of control subjects with normal glucose tolerance and preserved FPIS. Group 2 included patients with IGT and a reduced FPIS. Group 3 consisted of subjects with IGT but normal FPIS. Both groups were paired by age and BMI with subjects in the control group. Subjects underwent an intravenous glucose tolerance test to classify each case, and then a load with a mixed meal load to measure postprandial lipidemia. A total of 32 subjects were evaluated: 10 were control subjects, 8 subjects with IGT with a reduced FPIS and 14 subjects with IGT and preserved FPIS. After administration of a standardized meal, group 2 showed a greater glucose area under the curve (AUC) at 30 and 120 min (p=0.001, for both). This group also showed a statistically significant increase (p<0.001) in triglyceride AUC. A reduced FPIS is significantly and independently associated with a larger postprandial hyperlipidemia in subjects with IGT.


The effect of administering ketoprofen on the physiology and behavior of dairy cows following surgery to correct a left displaced abomasum.
Surgical correction of left displaced abomasum (LDA) is common in lactating dairy cattle. Despite the growing acceptance that abdominal surgery is painful, few cows are administered analgesia following LDA surgery. The objective of this research was to examine the effect of administering a label dose of ketoprofen on physiological and behavioral indicators of pain in dairy cattle. Holstein cows were enrolled in a field study following LDA surgery. Surgery was performed using the standing right flank (RF) approach or the paramedian (PARA) approach. Using a triple-blind randomized trial, each animal was assigned to receive either 3mg of ketoprofen/kg of body weight or saline (the equivalent volume) by intramuscular injection immediately following surgery and 24h postoperatively. Physiological parameters (heart rate, respiration rate, and rumen motility), blood β-hydroxybutyrate (BHBA) levels, and an assessment of cow attitude were measured on the day of surgery, and at 2 follow-up visits (visit 1=3 ± 0.9 d and visit 2=9 ± 1.2 d postsurgery; n=175). Milk production and culling were recorded for all cows enrolled in the study. Producers assessed their cows' attitudes and appetites daily for the first 3 d following surgery. A subset of cows (n=37) were fitted on the day of surgery with a 3-axis accelerometer on the hind leg to assess lying behavior. Continuous and binary outcome data were analyzed using multivariable mixed linear and mixed logistic models, respectively, with cow modeled as a random effect. Ketoprofen did not alter the physiological measures, BHBA levels, or behavioral outcomes measured. Cows subjected to RF surgery had longer lying times [model coefficient β=228.9 min; 95% confidence interval (CI): 122.2 to 335.6] in the first 3 d following surgery, and lower heart rates (β=-9.4 beats/min; 95% CI: -12 to -6.9 beats/min) at the follow-up visits, compared with animals that underwent PARA surgery. Regardless of surgical procedure, BHBA decreased from surgery day to visit 1 (β=-1.9 mmol/L; 95% CI: -2.1 to -1.7) and visit 2 (β=-2.0 mmol/L; 95% CI: -0.2.2 to -1.8). Producer reports indicated that animals that received ketoprofen were more likely to begin eating when provided fresh feed during the first 3 d following surgery compared with those that received saline (odds ratio=4.8; 95% CI: 0.97 to 23.8). These results indicate that PARA surgery may be more painful relative to lying down than the RF approach. The apparent differences in appetite or attitude in cows that received ketoprofen reported by producers warrant further investigation.


The Direct Healthcare Expenditures of Prostate Cancer by Disease Severity: Evidence from US National Survey Data.
Background: In the United States, approximately 2.8 million men have a history of prostate cancer (PC). Objective: This study quantified the effects of PC, overall and by disease severity on direct healthcare costs to insurers and patients. Methods: Using 1996-2010 data from the Medical Expenditure Panel Survey (MEPS), a large, nationally representative US database, multivariate analyses were used to assess the relationship between PC and direct annual healthcare costs to insurers and patients, at individual and US aggregate levels. Men aged 40 years and older with International Classification of Diseases, Ninth Revision (ICD-9) diagnosis code 185 were identified. Disease severity was determined with clinical assistance and based, in part, on the data in MEPS. The cohorts were: localized cancer not treated with chemotherapy, localized cancer treated with chemotherapy, and metastatic cancer. Results: The MEPS database included 1297 patients with PC: 811 patients with localized PC not treated with chemotherapy, 426 patients with PC treated with chemotherapy, and 60 patients with metastatic PC. PC had a larger effect on incremental costs for metastatic patients, $20 357, vs $16 709 for localized PC with chemotherapy, and $5238 for localized PC with no chemotherapy. When aggregated to the US population, PC accounted for an incremental annual cost of $15 billion. The largest aggregate annual costs were incurred by patients with localized PC treated with chemotherapy ($8.6 billion), compared to those not treated with chemotherapy ($4.8 billion) and metastatic patients ($1.6 billion). Conclusions: The aggregate annual costs of PC are substantial for all groups examined and greatest for patients with localized cancer treated with chemotherapy. This reflects the relatively high prevalence and high per capita healthcare expenditures associated with this group. With a growing and aging population, the prevalence of PC is expected to rise, increasing the burden on public health.


Postsynaptic alpha-adrenoceptor subtypes in rabbit blood vessels and rat anococcygeus muscle studied in vitro.
Postsynaptic alpha-adrenoceptor subtypes were investigated in vitro, employing rabbit aorta, pulmonary artery, and portal vein, and rat anococcygeus. Phenylephrine (alpha 1-selective), alpha-methylnoradrenaline (mixed agonist), and xylazine (alpha 2-selective) were used as agonists, and prazosin (alpha 1-selective) and rauwolscine (alpha 2-selective) as antagonists. In all tissues, agonist concentration-response curves were monophasic and their shape was unaltered by either antagonist. In rabbit blood vessels, prazosin was as potent against alpha-methylnoradrenaline as against phenylephrine and was 1,000 times more potent than rauwolscine; xylazine was a partial agonist of low potency. Hence, the postsynaptic receptors of these tissues are alpha 1. In the rat anococcygeus, there was some evidence for an alpha 2-receptor: xylazine was a full and potent agonist, rauwolscine was more potent against xylazine than against alpha-methylnoradrenaline or phenylephrine, and in experiments protecting against irreversible blockade by phenoxybenzamine, xylazine afforded significantly greater protection to alpha-methylnoradrenaline than to phenylephrine. However, prazosin was more potent than rauwolscine against xylazine and was equipotent against all agonists. Hence, although the smooth muscle cells of the rat anococcygeus may contain alpha 2-adrenoceptors, the predominant population is alpha 1.


Rupatadine protects against pulmonary fibrosis by attenuating PAF-mediated senescence in rodents.
A similar immune response is implicated in the pathogenesis of pulmonary fibrosis and allergic disorders. We investigated the potential therapeutic efficacy and mechanism of rupatadine, a dual antagonist of histamine and platelet-activation factor (PAF), in bleomycin- (BLM-) and silica-induced pulmonary fibrosis. The indicated dosages of rupatadine were administered in rodents with bleomycin or silica-induced pulmonary fibrosis. The tissue injury, fibrosis, inflammatory cells and cytokines, and lung function were examined to evaluate the therapeutic efficacy of rupatadine. The anti-fibrosis effect of rupatadine was compared with an H1 or PAF receptor antagonist, and efforts were made to reveal rupatadine's anti-fibrotic mechanism. Rupatadine promoted the resolution of pulmonary inflammation and fibrosis in a dose-dependent manner, as indicated by the reductions in inflammation score, collagen deposition and epithelial-mesenchymal transformation, and infiltration or expression of inflammatory cells or cytokines in the fibrotic lung tissue. Thus, rupatadine treatment improved the declined lung function and significantly decreased animal death. Moreover, rupatadine was able not only to attenuate silica-induced silicosis but also to produce a superior therapeutic efficacy compared to pirfenidone, histamine H1 antagonist loratadine, or PAF antagonist CV-3988. The anti-fibrotic action of rupatadine might relate to its attenuation of BLM- or PAF-induced premature senescence because rupatadine treatment protected against the in vivo and in vitro activation of the p53/p21-dependent senescence pathway. Our studies indicate that rupatadine promotes the resolution of pulmonary inflammation and fibrosis by attenuating the PAF-mediated senescence response. Rupatadine holds promise as a novel drug to treat the devastating disease of pulmonary fibrosis.


Should limb revascularization take priority over dissection repair in type a aortic dissection presenting as isolated acute limb ischaemia.
A best evidence topic in cardiac surgery was written according to a structured protocol. The question addressed was should limb revascularization take priority over dissection repair in acute type A aortic dissection (ATAAD) presenting as isolated acute limb ischaemia? Altogether 133 papers were found using the reported search, of which 7 represented the best evidence to answer the clinical question. The authors, journal, date and country of publication, patient group studied, study type, relevant outcomes and results of these papers are tabulated. Six studies showed that aortic repair alone resulted in the reperfusion of 60-100% of ischaemic lower limbs and recommended a strategy prioritizing aortic repair. In those with persistent isolated limb ischaemia post-aortic repair, expeditious peripheral revascularization procedures produced excellent patient outcomes comparable to those of ATAAD patients without malperfusion syndromes. In the remaining study, aortic repair was delayed in order to prioritize percutaneous reperfusion therapy aimed at treating the peripheral malperfusion. However, this resulted in one-third of patients not surviving to aortic surgery. We conclude that delaying aortic repair for limb reperfusion procedures results in unacceptably high mortality rates and that repair alone results in high reperfusion rates of ischaemic limbs. We therefore strongly recommend that aortic repair be the primary therapy for ATAAD patients even when limb ischaemia is the presenting feature. Limbs should be reassessed immediately after aortic repair and revascularization procedures undertaken urgently if any pulse deficits remain.


[Effect of Risperidone on BDNF-TrkB Signaling Pathway in Rat Brain].
To investigate the effect of risperidone on the expression of brain-derived neurotrophic factor (BDNF) and its receptors, tyrosine kinase receptor (TrkB) and P75 neurotrophin receptor (P75NTR) in rat brain. Sixteen SD rats were divided into two groups (n = 8 for each group). The rats in experimental group were treated with risperidone [0.25 mg/(kg · d)] for 14 d, while the control group was given placebo. Total RNA sample in prefrontal cortex, temporal cortex and hippocampus was extracted, and the expression of BDNF, TrkB and P75NTR mRNA were determined by quantitative real-time PCR. The treatment of risperidone significantly up-regulated the expressions of BDNF and TrkB in prefrontal cortex, temporal cortex and hippocampus, while the expression of P75NTR was not significantly changed. Risperidone upregulated BDNF-TrkB signaling, but not BDNF-P75NTR signaling, which may be helpful for the further pharmacological study of risperidone.


[Rearrangement of the RET gene in papillary thyroid carcinoma].
The RET/PTC oncogenes, activated forms of the RET protooncogene, almost exclusively found in papillary thyroid carcinoma (PTC). What is more, the targeted expression of RET/PTC in mice leads to the development of thyroid tumors very similar to human PTCs. In all RET/PTC types the RET tyrosine kinase domain is fused to the N-terminus of ubiquitously expressed genes that is capable of ligand-independent dimerization. The majority of RET/PTC identified consists of two types which results from the inversion of chromosome 10: RET/PTC1 and RET/PTC3. The prevalence of RET/PTC in papillary thyroid carcinomas of thyroid varies widely from a few to about 80% with the highest frequency in tumors arising in children after ionizing radiation. In Polish population the frequency of RET rearrangements in papillary cancers is 27%, although, it was reported to be twice higher in young patients (50% in patients younger than 21 at operation). Correlation with clinical outcome as well as prognostic value of RET/PTC is controversial. Some authors suggest that it predicts metastases, others found rearranged RET in more favourable, slow growing tumors. RET/PTC3 seems to be associated with solid/follicular variant PTC and short latency period (it is found more frequently in children) whereas RET/PTC1--with classic PTC variant and long latency.


Circumventricular Organ Apelin Receptor Knockdown Decreases Blood Pressure and Sympathetic Drive Responses in the Spontaneously Hypertensive Rat.
The central site(s) mediating the cardiovascular actions of the apelin-apelin receptor (APJ) system remains a major question. We hypothesized that the sensory circumventricular organs (CVOs), interfacing between the circulation and deeper brain structures, are sites where circulating apelin acts as a signal in the central nervous system to decrease blood pressure (BP). We show that APJ gene (aplnr) expression was elevated in the CVOs of spontaneously hypertensive rats (SHRs) compared to normotensive Wistar Kyoto (WKY) controls, and that there was a greater mean arterial BP (MABP) decrease following microinjection of [Pyr1]apelin-13 to the CVOs of SHRs compared to WKY rats. Lentiviral APJ-specific-shRNA (LV-APJ-shRNA) was used to knockdown aplnr expression, both collectively in three CVOs and discretely in individual CVOs, of rats implanted with radiotelemeters to measure arterial pressure. LV-APJ-shRNA-injection decreased aplnr expression in the CVOs and abolished MABP responses to microinjection of [Pyr1]apelin-13. Chronic knockdown of aplnr in any of the CVOs, collectively or individually, did not affect basal MABP in SHR or WKY rats. Moreover, knockdown of aplnr in any of the CVOs individually did not affect the depressor response to systemic [Pyr1]apelin-13. By contrast, multiple knockdown of aplnr in the three CVOs reduced acute cardiovascular responses to peripheral [Pyr1]apelin-13 administration in SHR but not WKY rats. These results suggest that endogenous APJ activity in the CVOs has no effect on basal BP but that functional APJ in the CVOs is required for an intact cardiovascular response to peripherally administered apelin in the SHR.


[Ionizing radiation as a risk factor for thyroid cancer in Krakow and Nowy Sacz regions].
In a group of 224 patients with thyroid cancer registered from 1990 to 1999 and in 243 controls matched for sex and age doses for the thyroid due to accumulation of 131I after Chernobyl accident and radiological diagnostic procedures involving the neck and dentition was calculated according to Johnson's model and based upon questionnaires. There was a significantly higher dose for the thyroid due to accumulation of 131I in the Krakow area in comparison with the Nowy Sacz area. There was also a significantly higher frequency of radiological procedures involving the thyroid gland preceding the development of thyroid cancer in comparison with the control group. The present findings are concordant with about 40% increase of thyroid cancer in the Krakow area in comparison with the Nowy Sacz region. The present study has indicated that ionising radiation may be a factor involved in the pathogenesis of thyroid cancer and its significant rise within the last 10 years. However, it has not been the main factor, but only one of the risk factors. The study has also indicated that iodine prophylaxis should be continued and that radiological procedures involving the neck and head in patients with nodular goitre should be carried out with caution.


Influence of formulation variables on the in-vitro release of albumin from biodegradable microparticulate systems.
Poly(D,L-lactide-co-glycolide) microspheres containing BSA were prepared by a modified solvent evaporation method using a double emulsion. These microspheres were characterized for size, morphology, surface absorbed protein, encapsulation efficiency and release kinetics. The influence of two formulation variables (the procedure to obtain the first emulsions and the lyophilization of the microspheres once obtained) on the physical characteristics and release behaviour of the microspheres was also investigated. Sonicated microspheres were smooth and spherical, with a mean particle size of 20 microns and an encapsulation efficiency of 81%. When the first emulsion was prepared by vortex mixing the particles were irregular and porous, with a mean size of 31 microns and a lower encapsulation efficiency (56%). The sonication allows a more homogeneous emulsion as well as a lower percentage of albumin adsorbed on the surface. The in vitro release profile was described as a biexponential process with an initial burst effect due to the release of the protein adsorbed on the microsphere surface and a second sustained release phase due to protein diffusion through the channels or pores formed in the polymer coat. The release of BSA was dependent on the preparation method. The greatest burst release was found for microspheres formulated using the vortex mixer, 58% of the encapsulated protein was released during the first 24 h, whereas sonicated microspheres released 32.2%. This burst effect could be reduced by lyophilizing the microspheres following their preparation. The amount of protein released decreased to 28.3% and 51.6% in sonicated and non-sonicated microspheres respectively, when they were lyophilized.


[Detection of the antigenic peptides in pHLA complexes by ultrafiltration-high performance liquid chromatography].
Recombinant HLA-Ⅰ molecules/antigenic peptide complexes (pHLA complexes) are applied in the research of human T cell-specific immune responses. The preparation of pHLA complex is based on genetic engineering and protein in vitro dilution and folding-refolding technology. In an in vitro refolding system, recombinant HLA-Ⅰ molecules correctly fold and bind with antigenic peptides to form complexes. In this study, ultrafiltration-high performance liquid chromatography (ultrafiltration-HPLC) was used for quantitative determination of the antigenic peptides in recombinant pHLA complexes, especially for those in a small amount of prepared products. By adding the recombinant HLA-Ⅰ molecules and antigenic peptides into the refolding buffer, the heavy chain (HC) and light chain (β2m) of recombinant HLA-Ⅰ molecules were refolded and bond with the VYF antigenic peptide containing anchor residues to form a pHLA complex. The unbound free antigenic peptide VYF was removed by ultrafiltration to retain the complex. Finally, the pHLA complex was treated by acid to destroy its interaction, thus releasing the antigenic peptide. The results showed that the prepared recombinant pHLA complex was recognized by HLA-Ⅰ molecule specific antibody W6/32, which indicated that the recombinant HLA-Ⅰ class molecule had correct folding and was identified as pHLA complex. The antigen peptide VYF contained in the pHLA complex was also detected by ultrafiltration-HPLC, so it is feasible to apply ultrafiltration-HPLC for determination of pHLA complex. Compared with Western blotting, the concentration of antigenic peptides detected by ultrafiltration-HPLC was 0-9 μg/mL. The binding conditions can be optimized according to the amount of antigenic peptides bound in the complex in order to improve the folding efficiency of HLA-Ⅰ molecules and promote the binding of HLA-Ⅰ molecules to antigenic peptides. The production rate of pHLA complexes in the refolding system can also be calculated according to the content of antigenic peptides bound by pHLA complexes. Therefore, ultrafiltration-HPLC in this study can be used for the quality control of the preparation process of pHLA complexes, and may facilitate the research of T cell-specific immunity, artificial antigen-presenting cells, and development of specific tetramer probe applications.


Metabolic activation in the fetal mouse salivary gland culture system with rat hepatocytes, rat S-9, and human S-9.
The usefulness of an in vitro assay for embryotoxicity may depend on the availability of metabolic activation systems that will function in the culture system. The fetal mouse salivary gland has been investigated as an in vitro assay system. To see if the glands would grow in the presence of metabolic activators and if the glands would react to metabolites known to be embryotoxic, the glands were grown in the presence of cyclophosphamide (CP) and several activation systems. These included isolated rat hepatocytes, uninduced rat S-9, rat S-9 induced with 3-methylcholanthrene (3-MC), rat S-9 induced with Aroclor 1254, and human S-9. Twenty salivary glands were isolated from 13 day embryos (plug day = 0) and were grown in each treatment for 48 h. One control had no activation system of CP, one had an activation system but no CP, and three treatments had the activation system and 25, 75, or 150 micrograms/ml CP. The S-9 with cofactors and the appropriate amount of CP was contained in dialysis bags. The greatest suppression of salivary gland growth occurred in co-culture with hepatocytes activating CP. The S-9 induced by Aroclor 1254 was nearly as effective as the hepatocytes. The next most effective was a group with similar activity consisting of the uninduced rat S-9 and the three samples of human S-9. The 3-MC-induced S-9 was the least effective in suppressing growth of salivary glands. All the activation systems tested can be used with the salivary gland culture system.


Insulin induction of pip 92, CL-6, and novel mRNAs in rat hepatoma cells.
Insulin directly affects many aspects of cellular metabolism. An additional, poorly studied effect of insulin is the regulation of multiple genes whose products are important in many cellular functions. Using differential screening techniques, we identified insulin-regulated genes induced in insulin-treated rat H4IIE (H4) hepatoma cells. Two of the mRNAs identified were homologous to the previously characterized mouse pip 92 and rat CL-6 immediate-early genes. The pip 92 clone was initially isolated from mitogen-stimulated mouse Balb/c 3T3 fibroblasts, whereas the CL-6 clone was first isolated from regenerating rat liver. In this article, we demonstrated that in rat H4 cells, the transcription rates of both pip 92 and CL-6 are induced by insulin alone. Additionally, we showed that the transcription rates of two other genes, whose sequences are not homologous to any other sequences in gene bank ("novel genes"), were rapidly and transiently induced by insulin. These results demonstrate that insulin regulates the expression of several novel genes with a time-course similar to members of the immediate-early response gene family.


The surgical management of traumatic c6-c7 spondyloptosis.
A case of traumatic spondyloptosis of the cervical spine at the C6-C7 level is reported. The patient was treated succesfully with a anterior-posterior combined approach and decompression. The patient had good neurological outcome after surgery. A-51-year-old female patient was transported to our hospital's emergency department after a vehicle accident. The patient was quadriparetic (Asia D, MRC power 4/5) with severe neck pain. Plain radiographs, computerize tomography and spinal magnetic resonance imaging (MRI) showed C6-7 spondyloptosis and C5, C6 posterior element fractures. Gardner-Wells skeleton traction was applied. Spinal alignment was reachived by traction and dislocation was decreased to a grade 1 spondylolisthesis. Then the patient was firstly operated by anterior approach. Anterior stabilization and fusion was firstly achieved. Seven days after first operation the patient was operated by a posterior approach. The posterior stabilization and fusion was achieved. Postoperative lateral X-rays and three-dimensional computed tomography showed the physiological realignment and the correct screw placements. The patient's quadriparesis was improved significantly. Subaxial cervical spondyloptosis is a relatively rare clinical entity. In this report we present a summary of the clinical presentation, the surgical technique and outcome of this rarely seen spinal disorder.


Metabolomics in endometriosis: challenges and perspectives for future studies.
Endometriosis is a complex disease characterized by inflammation and the growth of endometrial- like glands and stroma outside the uterine cavity. The pathophysiology of endometriosis is not entirely understood, however, with a prevalence of ~10% of women in their reproductive years, the disease symptoms significantly affect the quality of life of millions of women globally. Metabolomic studies have previously identified specific metabolites that could be a signature of endometriosis. This approach could potentially be used as a non-invasive tool for early diagnosis and provide a better understanding of endometriosis pathophysiology. This review aims to provide insight as to how endometriosis affects the metabolome by reviewing different studies that have used this approach to design follow-up studies. The search query included the term 'endometriosis' in combination with 'metabolomics', 'lipidomics', or 'sphingolipidomics' published between 2012 and 2020. We included studies in humans and animal models. Most studies reported differences in the metabolome of subjects with endometriosis in comparison to healthy controls and used samples taken from serum, endometrial tissue, follicular fluid, urine, peritoneal fluid, or endometrial fluid. Statistically significant metabolites contributed to group separation between patients and healthy controls. Reported metabolites included amino acids, lipids, organic acids, and other organic compounds. Differences in methods, analytical techniques, and the presence of confounding factors can interfere with results and interpretation of data. Metabolomics seems to be a promising tool for identifying significant metabolites in patients with endometriosis. Nonetheless, more investigation is needed in order to understand the significance of the study results. Endometriosis is a chronic disease affecting the quality of life in one out of every ten women during their reproductive years, causing pain and infertility. It is characterized by inflammation and growth of tissue like the endometrium (uterus lining) outside the uterine cavity. Studies have searched for a predictor of endometriosis-associated changes by observing small molecules necessary for metabolism on a large scale (metabolomics). Metabolomics could serve to resolve one of the biggest challenges that patients with endometriosis face: a delay in diagnosis. In this review, the authors summarize identified potential biomarkers from various bodily fluids and tissues that are characteristic of metabolic processes observed in endometriosis. Biomarkers include cell growth, cell survival, high energy demand, oxidative stress, and fatty acid levels. A metabolomics approach offers promise as a non-invasive tool to identify significant metabolite changes in patients with endometriosis, potentially leading to earlier diagnoses and new opportunities for back-translational strategies.