Industrial lab-on-a-chip: design, applications and scale-up for drug discovery and delivery.
Microfluidics is an emerging and promising interdisciplinary technology which offers powerful platforms for precise production of novel functional materials (e.g., emulsion droplets, microcapsules, and nanoparticles as drug delivery vehicles- and drug molecules) as well as high-throughput analyses (e.g., bioassays, detection, and diagnostics). In particular, multiphase microfluidics is a rapidly growing technology and has beneficial applications in various fields including biomedicals, chemicals, and foods. In this review, we first describe the fundamentals and latest developments in multiphase microfluidics for producing biocompatible materials that are precisely controlled in size, shape, internal morphology and composition. We next describe some microfluidic applications that synthesize drug molecules, handle biological substances and biological units, and imitate biological organs. We also highlight and discuss design, applications and scale up of droplet- and flow-based microfluidic devices used for drug discovery and delivery.


Hydrocephalus despite extra ventricular drainage in adults: a new description of multiloculated hydrocephalus.
Multiloculated hydrocephalus constitutes a challenging pathology due to intracerebral haemorrhage or ventriculitis leading to iterative shunt revision frequently described in paediatric neurosurgery, but poorly reported in adults. Nevertheless, this potential complication of intraventricular haemorrhage, already drained in emergency, should be considered with special interest, as ideal management of cerebrospinal drainage remains debated in such situation. We thus report herein the case of intraventricular haemorrhage in an adult complicated of multiloculated hydrocephalus, as an illustrative plea for endoscopic surgery.


Schizophrenia psychopathology in a Kuwaiti Arab sample.
The heterogeneity of schizophrenia psychopathology has led to a search for symptom clusters that could be related to broad features of the disease. The objectives of the study were to: (1) highlight the pattern of symptoms among Kuwaiti subjects with schizophrenia, using the ICD-10 symptom checklist; (2) assess the factor structure of the Brief Psychiatric Rating Scale 18 (BPRS-18) and BPRS-24, and (3) evaluate the relationship of the resulting factors with sociodemographic characteristics, age at onset of illness, family history of mental illness, objective indicators of quality of life, self-esteem, affect balance, family caregiver burden, patient's perceptions of service satisfaction and needs for care. Consecutive outpatients in stable condition were assessed with the BPRS and ICD-10 symptom checklist, as well as measures of quality of life, needs for care, service satisfaction and family caregiver burden. There were 130 patients (66.1% men, mean age = 36.8 years, age at onset of illness = 24.2 years, duration of illness = 12.9 years). Of the ICD-10 symptoms, the commonest positive symptoms were hallucinations (58.5%) and delusions (72%). Catatonic symptoms were rare (2.9%). About a quarter of the subjects experienced 4 of the negative symptoms. In exploratory factor analysis, we broadly replicated the known syndromes for BPRS-24 (disorganization, positive, activation, manic, negative and depression) and BPRS-18 (negative, positive, activation and affect). In regression analyses, the variables independently associated with psychopathology were family income, negative affect, self-esteem, duration of illness, age, lack of money for enjoyment, met needs for care and caregiver tension. The negative syndrome had more significant associations with the variables investigated than the positive syndrome. Psychopathological scores were separable from indices of psychological well-being. In this first study of schizophrenia psychopathology from the Arab world, the clinical manifestations were similar to the data from the developed countries. The persistence of psychotic symptoms despite freely available antipsychotic treatment and the impact on caregiver burden call for attention to the perennial issue of treatment resistance and underscore the need for continued interaction with family members after the acute inpatient phase of treatment, in order to address the impact of symptoms.


Carry-over of melamine from feed to eggs and body tissues of laying hens.
The objective of this study was to assess the carry-over of melamine from feed into eggs and body tissues of laying hens. In the first experiment, laying hens were supplied with feed added at 0, 1, 2, 5, 25, 50, and 100 mg kg(-1) of melamine for 21 days followed by a depletion period to observe the residues of melamine in eggs. In a second experiment, laying hens were allocated 0, 50, and 100 mg kg(-1) melamine to determine levels of melamine in body tissues. Melamine and cyanuric acid were simultaneously analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the diet as well as in eggs and body tissue. In the first experiment, melamine appeared in the egg within 24 h after first ingestion of the melamine at 5, 25, 50, and 100 mg kg(-1). Melamine concentration in egg reached a maximum of 2.34 mg kg(-1) within 17 days after exposure of 100 mg kg(-1) melamine, and the carry-over rate for melamine from feed to the eggs was 1.21%. In the second experiment, melamine was detected in tissues within 3 days after exposure; the maximum concentration of melamine residues occurred in the 100 mg kg(-1) group and was as follows: egg (1.83) > kidney (1.21) > breast muscle (0.86) > liver (0.70) > serum (0.42). The melamine level in egg albumen was about twice that of egg yolk. Melamine levels in laying hens decreased rapidly with withdrawal from feed, but melamine only declined to undetectable levels in the egg at day 6 and in tissues at day 4 after last ingestion of 100 mg kg(-1). It can be concluded that a pathway exists for the transmission of melamine from feed to egg and body tissues and the carry-over rate of melamine is low, and that melamine is not metabolized into cyanuric acid in laying hens. A positive relationship exists between exposure levels and eggs or tissues, but no direct relationship between the exposure time and measured levels of melamine in eggs and tissues. The current Chinese limit for melamine in feed and feed material of 2.5 mg kg(-1) established on 8 June 2009 is sufficient to ensure that levels in eggs, breast muscle, liver as well as kidney will not exceed the limit of 2.5 mg kg(-1) in foods other than infant formula introduced by many countries including China.


Highly reproducible bactericidal activity test results by using a modified National Committee for Clinical Laboratory Standards broth macrodilution technique.
Bactericidal testing historically has exhibited variable reproducibility, even when prior standardized methods were employed. Several modifications to the National Committee for Clinical Laboratory Standards (NCCLS) broth macrodilution method are proposed to improve reproducibility. Recommended changes from the approved NCCLS guidelines (M21-A and M26-A) include omitting serum supplementation of Mueller-Hinton broth, incubating tubes at 35 degrees C for 24 h with no agitation until they are sampled, running all tests in duplicate with six dilutions instead of nine, reincubating the test for an additional 24 h to resolve discrepant bactericidal activity test results, using a single 0.1-ml sample from each clear tube for subculture, and adopting an alternate method for calculating endpoint determination. In order to test these recommendations in a clinical laboratory setting, we used the modified methodology on 224 separate tests for bactericidal activity. There were 102 serum bactericidal titer (SBT) and 122 minimum bactericidal concentration (MBC) assays performed. By defining reproducibility as agreement between duplicate tests +/- 1 dilution, we found 207 of 224 tests (92%) were reproducible at the 24-h subculture point (94% for the SBT assay and 91% for the MBC assay). When the 17 assays with discrepant results were incubated an additional 24 h for a second subculture, only 1 of 224 tests (0.4%) remained discrepant. The method used is practical for a clinical laboratory that chooses to perform bactericidal activity testing and assures a high level of reproducibility between duplicate assays. The total cost of a test was approximately $25.00.


Infection with hepatitis viruses, FIB-4 index and risk of hepatocellular carcinoma in southern Italy: a population-based cohort study.
The incidence of hepatocellular carcinoma (HCC) and its association with hepatitis C (HCV) and hepatitis B virus (HBV) infections, FIB-4 index and liver enzymes was assessed in an area of the province of Naples covered by a population-based cancer registry. We conducted a cohort investigation on 4492 individuals previously enrolled in a population-based seroprevalent survey on HCV and HBV infections. The diagnosis of HCC was assessed through a record linkage with the cancer registry. Hepatic metabolic activity was measured through serum alanine transaminase, aspartate aminotransferase, gamma-glutamyl-transferase, and platelet. The FIB-4 index was used as a marker of fibrosis. We computed HCC incidence rates (IR) for 100,000 (105) person-years of observation, and multivariable hazard ratios (HR) with 95 % confidence intervals (CI) to assess risk factors for HCC. Twenty two cases of HCC were diagnosed during follow-up (IR = 63.3 cases/105). Significantly increased HCC risks were documented in individuals with higher than normal liver enzymes and low platelet count; in the 239 HCV RNA-positives (HR = 61.8, 95 % CI:13.3-286); and in the 95 HBsAg-positives (HR = 75.0) -as compared to uninfected individuals. The highest FIB-4 score was associated with a 17.6-fold increased HCC risk. An elevated FIB-4 index turned out to be an important predictor of HCC occurrence. Although the standard method to assess hepatic fibrosis in chronic hepatitis remains the histologic staging of liver biopsy specimen, the assessment of FIB-4 in HCV RNA-positive individuals may help in identifying the highest HCC-risk individuals who need anti-HCV treatment most urgently.


The Biplanar Effect of the Medial Cuneiform Osteotomy.
Background. The "foot-CORA" (center of rotation of angulation) method confirms the medial cuneiform as the site of deformity in most forefoot/midfoot deformities and is therefore the ideal location to correct those deformities. It has been consistently observed intraoperatively by the senior author that there is a secondary, unintentional deformity created in the transverse plane when dorsiflexion and plantar flexion osteotomies of the medial cuneiform are performed to correct pronation and supination forefoot deformities, respectively. These effects may not be desirable. This biplanar effect of medial cuneiform osteotomies has been observed but not studied. The purpose of this study was to perform the 4 commonly used medial cuneiform osteotomy techniques on cadaveric feet to demonstrate their biplanar effects. Methods. Four formaldehyde preserved cadaveric feet were used to perform 4 techniques of medial cuneiform osteotomy: dorsiflexion plantar-based opening wedge, plantar flexion dorsal-based opening wedge, dorsiflexion dorsal-based closing wedge, and plantar flexion plantar-based closing wedge. Photographs and fluoroscopy were used to assess the angular changes in the sagittal and transverse planes. Angular measurements were made using OsiriX software on fluoroscopic images. Results. The medial cuneiform opening wedge osteotomies produced midfoot abduction in addition to the desired dorsiflexion and plantar flexion. The medial cuneiform closing wedge osteotomies produced midfoot adduction in addition to the desired dorsiflexion and plantar flexion. Conclusion. We confirm that intentional sagittal uniplanar osteotomies of the medial cuneiform create obligate biplanar effects. This is likely a result of tethering by ligaments and the joint capsules on the lateral border of the medial cuneiform. The obligate transverse plane effect can be used to one's advantage or result in an undesired effect if not considered during surgical planning and execution. We propose a simple treatment algorithm for selecting the appropriate medial cuneiform osteotomy for forefoot/midfoot deformities. Levels of Evidence: Level V.


Radiographic localization of unerupted teeth in the anterior part of the maxilla: a survey of methods currently employed.
The radiographic prescribing habits of consultants in orthodontics and oral surgery are described with specific reference to assessment of unerupted maxillary canine teeth. Orthodontists use more radiographs than oral surgeons under these circumstances. The relative radiation for commonly used combinations of radiographs has been summarized and recommendations made regarding the use of certain of these.


Expanded analysis of secondary germline findings from matched tumor/normal sequencing identifies additional clinically significant mutations.
Next-generation sequencing (NGS) for tumor molecular profiling can reveal secondary germline pathogenic and likely pathogenic variants (LPV/PV). The American College of Medical Genetics (ACMG) recommends return of secondary results for a subset of 59 genes, but other genes with evidence of clinical utility are emerging. We previously reported that 4.3% of patients who underwent NGS of a targeted panel of 201 genes had LPV/PV based on the ACMG list. Here we report the frequency of additional germline cancer-related gene variants and discuss their clinical utility. Matched tumor and germline DNA NGS of a targeted panel of 201 genes was performed in a research laboratory on samples from 1000 patients with advanced or metastatic solid tumors enrolled in a molecular testing protocol (NCT01772771). The frequency of germline LPV/PV in 54 cancer-related genes, beyond the genes in ACMG list, were analyzed. Among 1000 patients who underwent tumor/normal DNA sequencing, 46 (4.6%) were found to have a germline LPV/PV in the following genes: AR-(5), ATM-(4), BAP1-(1), CDH1-(1), CDKN2A-(1), CHEK1-(2), CHEK2-(10), EGFR-(1), ERCC3-(4), ERCC5-(1), HNF1B-(1), HRAS-(1), MITF-(4), MLL3-(1), NF1-(3), PKHD1-(4), PTCH1-(1), and SMARCA4-(1). Thus, a total 8.7% of patients had an LPV/PV with 2 patients having 2 concomitant germline LPV/PV. Five mutations in high-penetrance hereditary cancer predisposition genes were selected to be returned to patients or their representatives: BAP1, CDH1, CDKN2A, EGFR, and SMARCA4. Broader genomic testing is likely to identify additional secondary pathogenic germline alterations, some with potential clinical utility for return to patients and their relatives. The recommended genes for which germline results should be returned are continually changing, warranting continued study.


Glucocorticoid/Adiponectin Axis Mediates Full Activation of Cold-Induced Beige Fat Thermogenesis.
Glucocorticoids (GCs), a class of corticosteroids produced by the adrenal cortex in response to stress, exert obesity-promoting effects. Although adaptive thermogenesis has been considered an effective approach to counteract obesity, whether GCs play a role in regulating cold stress-induced thermogenesis remains incompletely understood. Here, we show that the circulating levels of stress hormone corticosterone (GC in rodents) were significantly elevated, whereas the levels of adiponectin, an adipokine that was linked to cold-induced adaptive thermogenesis, were decreased 48 h post cold exposure. The administration of a glucocorticoid hydrocortisone downregulated adiponectin protein and mRNA levels in both WAT and white adipocytes, and upregulated thermogenic gene expression in inguinal fat. In contrast, mifepristone, a glucocorticoid receptor antagonist, enhanced adiponectin expression and suppressed energy expenditure in vivo. Mechanistically, hydrocortisone suppressed adiponectin expression by antagonizing PPARγ in differentiated 3T3-L1 adipocytes. Ultimately, adiponectin deficiency restored mifepristone-decreased oxygen consumption and suppressed the expression of thermogenic genes in inguinal fat. Taken together, our study reveals that the GCs/adiponectin axis is a key regulator of beige fat thermogenesis in response to acute cold stress.


Prevalence of impostor phenomenon and burnout in a Singapore health system.
Impostor phenomenon (IP) is a set of feelings encountered by individuals of being incompetent, despite experiencing successes. IP affects not only individuals on a personal level, but also organisations where the leadership diversity decreases due to employees' self-doubt. We aim to investigate the prevalence of IP and burnout among employees in the National University Health System (NUHS). All permanently employed full-time NUHS employees aged 21 years and above were invited to participate in this self-administered cross-sectional study between April 2021 and August 2021. Mass emails with the embedded study link were sent every 2-3 weeks to the employees' corporate email accounts. In our study, 61% of our study respondents reported having IP experiences and 97% reported having burnout. The associations of IP with ethnicity and age group were significant. Post hoc tests, however, showed that the association was statistically significant only in the 21-29 years age group. We found that there was no statistical significance between gender and the Maslach Burnout Inventory (MBI) profile types. However, we found that IP was significantly associated with individuals in the 21-29 years age group. This could be because younger individuals who just entered workforce may feel uncomfortable with their newfound independence and responsibility. Workplace support, such as workshops, and emotional support were found to be useful in helping individuals cope with IP. Future studies could be done post coronavirus disease 2019 (COVID-19) pandemic among healthcare workers to have a larger sample size to determine true IP and burnout prevalence.


Inhibitors of CDP-choline synthesis, action potential calcium channels, and stimulus-secretion coupling.
The effects of putative transmethylation inhibitors were tested on stimulus-secretion coupling and neurotransmitter secretion at synapses between neuroblastoma X glioma hybrid cells and myotubes. 5'-Deoxy-5'-isobutylthio-3-deazaadenosine or 5'-deoxy-5'-isobutylthioadenosine inhibited CDP-choline synthesis catalyzed by cholinephosphate cytidylyltransferase (CTP:cholinephosphate cytidylyltransferase, EC 18.104.22.168) and thereby decreased the rate of phosphatidylcholine synthesis from CDP-choline, but did not affect the transmethylation pathway for phosphatidylcholine synthesis. These compounds also inhibited 45Ca2+ uptake by hybrid cells mediated by voltage-sensitive Ca2+ channels, acetylcholine secretion at synapses, and signal transduction through cell membranes mediated by myotube nicotinic acetylcholine receptors. In contrast, 3-deazaadenosine or adenosine inhibited the transmethylation pathway for phosphatidylcholine synthesis, but had no effect on Ca2+ action potentials, acetylcholine secretion, or signal transduction through cell membranes mediated by nicotinic acetylcholine receptors. These results show that the stimulus-secretion coupling and secretion reactions studied are not dependent on phospholipid methylation and suggest that the activity of action potential Ca2+ channels and the rate of neurotransmitter secretion are functionally coupled to the rate of phosphatidylcholine synthesis via the CDP-choline pathway.


Measurement of serum levels of macrophage colony-stimulating factor (M-CSF) in patients with uremia.
Serum levels of monokines, including macrophage colony-stimulating factor (M-CSF), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1 alpha) and IL-1 beta (IL-1 beta), were measured in patients with chronic renal failure in an attempt to clarify the kinetics of these cytokines in the course of renal anemia. M-CSF was the only monokine detectable in the serum from all patients as well as healthy donors, making this cytokine feasible and reliable for serial evaluations. On all occasions, the level of M-CSF in uremic patients was significantly higher than that in healthy donors (29.4 +/- 12.3 vs. 5.5 +/- 1.1 ng/mL). In patients undergoing hemodialysis, the serum level of M-CSF was greater than that in patients undergoing continuous ambulatory peritoneal dialysis or in uremic patients without dialysis therapy. No difference was observed, however, in the levels of IL-1 alpha, IL-1 beta and TNF-alpha levels in these groups. Patients with severe anemia were subsequently treated with 60 to 80 U/kg per week of human recombinant erythropoietin (rhEpo) for 3 months. After this replacement therapy, hemoglobin levels increased with a variable change ranging from 0 to 3.5 g/dL. The pretherapy M-CSF level, however, was found to predict statistically the response to the therapy (p < 0.05). Patients with a lower pretherapy value responded better to rhEpo therapy; those with a higher level showed a minor degree of response. From these results, we postulate that the elevated M-CSF serum level in uremic patients is in part a consequence of the dialysis procedure and that rhEpo therapy is more effective in patients who are under sophisticated dialysis protocol and have a lower M-CSF level.


A Simple Pre-Operative Nuclear Classification Score (SPONCS) for Grading Cataract Hardness in Clinical Studies.
The aim of this study was to evaluate whether a simplified pre-operative nuclear classification score (SPONCS) was valid, both for clinical trials and real-world settings. Cataract classification was based on posterior nuclear color: 0 (clear), 1 (subcapsular/posterior cataract with clear nucleus), 2 (mild "green nucleus" with plus sign for yellow reflection of the posterior cortex), 3 (medium "yellow nucleus" with plus sign for brown/red posterior cortex reflection), 4 (advanced with 4 being "red/brown nucleus" and 4+ white nucleus), and 5 (hypermature/Morgagnian nucleus). Inter- and intra-observer validity was assessed by 30 Ophthalmologists for 15 cataract cases. The reliability of the cataract grading score in a surgical setting was evaluated. Correlation of nuclear scores was compared with phacoemulsification cumulative dissipated energy (CDE) in 596 patients. Analysis of mean intra-observer Cohen kappa agreement was 0.55 with an inter-observer score of 0.54 for the first assessment and 0.49 for the repeat assessment one week later. When evaluating results by nuclear color alone, there was a substantial agreement for both the intra-observer (0.70) and inter-observer parameters: 0.70 for the first test, and 0.66 on repetition with randomization of the cases after a week. CDE levels were found to be significantly different between all SPONCS score groups (p < 0.001), with a lower CDE related to a lower SPONCS score. A strong correlation was found between the SPONCS score and CDE (Spearman's rho = 0.8, p < 0.001). This method of grading cataract hardness is both simple and repeatable. This system can be easily incorporated in randomized controlled trials to lower bias and confounding effects regarding nuclear density along with application in the clinical setting.


Relations between the expression of p53, c-erbB-2, Ki-67 and HPV infection in cervical carcinomas and cervical dysplasias.
The analysis of mutual relations between HPV infection and the expression of cancer gene products and proliferative activity in cervical carcinomas and dysplasias. The expression of p53, c-erbB-2 oncoproteins and proliferative activity (Ki-67) was evaluated immunohistochemically in 41 cervical carcinomas and 29 dysplasias. HPV infection (type 16, 18) was assessed by in situ hybridization technique. HPV-positive carcinomas were found in 68.3% of cases. HPV type 16 infection were detected in 54% and HPV 18 in 39% of carcinomas. Simultaneous appearance of both virus types was shown in 25% of carcinomas. In dysplastic lesions, HPV infection was observed in 62.1% of cases. HPV type 16 was found in 34.5% and HPV 18 in 44.8% of patients. Both virus types were found in 17.2% of dysplasias. HPV infection was more extensive in cervical carcinomas than in dysplasias. Similarly the expression of oncoproteins was more intensive and referred to a higher percentage of cells in carcinomas. No relations between p53, c-erbB-2 overexpression and HPV infection were found. Ki-67 activity was found in a higher percentage of HPV-positive than in HPV-negative, both carcinomas and dysplasias. HPV infection, especially accompanied by increase of proliferative activity in dysplasias may define the cell subpopulation predisposed to malignant process development. The employment of in situ hybridization technique appears to be useful in detecting the viral infection in cytological smears even with no morphological changes in the cells.


In vivo dynamic compression has less detrimental effect than static compression on newly formed bone of a rat caudal vertebra.
Fusionless devices are currently designed to treat spinal deformities such as scoliosis by the application of a controlled mechanical loading. Growth modulation by dynamic compression was shown to preserve soft tissues. The objective of this in vivo study was to characterize the effect of static vs. dynamic loading on the bone formed during growth modulation. Controlled compression was applied during 15 days on the 7(th) caudal vertebra (Cd7) of rats during growth spurt. The load was sustained in the "static" group and sinusoidally oscillating in the "dynamic" group. The effect of surgery and of the device was investigated using control and sham (operated on but no load applied) groups. A high resolution CT-scan of Cd7 was acquired at days 2, 8 and 15 of compression. Growth rates, histomorphometric parameters and mineral density of the newly formed bone were quantified and compared. Static and dynamic loadings significantly reduced the growth rate by 20% compared to the sham group. Dynamic loading preserved newly formed bone histomorphometry and mineral density whereas static loading induced thicker (+31%) and more mineralized (+12%) trabeculae. A significant sham effect was observed. Growth modulation by dynamic compression constitutes a promising way to develop new treatment for skeletal deformities.


Intrinsic defect in T cell production of interleukin (IL)-13 in the absence of both IL-5 and eotaxin precludes the development of eosinophilia and airways hyperreactivity in experimental asthma.
Interleukin (IL)-5 and IL-13 are thought to play key roles in the pathogenesis of asthma. Although both cytokines use eotaxin to regulate eosinophilia, IL-13 is thought to operate a separate pathway to IL-5 to induce airways hyperreactivity (AHR) in the allergic lung. However, identification of the key pathway(s) used by IL-5 and IL-13 in the disease process is confounded by the failure of anti-IL-5 or anti-IL-13 treatments to completely inhibit the accumulation of eosinophils in lung tissue. By using mice deficient in both IL-5 and eotaxin (IL-5/eotaxin(-/-)) we have abolished tissue eosinophilia and the induction of AHR in the allergic lung. Notably, in mice deficient in IL-5/eotaxin the ability of CD4(+) T helper cell (Th)2 lymphocytes to produce IL-13, a critical regulator of airways smooth muscle constriction and obstruction, was significantly impaired. Moreover, the transfer of eosinophils to IL-5/eotaxin(-/-) mice overcame the intrinsic defect in T cell IL-13 production. Thus, factors produced by eosinophils may either directly or indirectly modulate the production of IL-13 during Th2 cell development. Our data show that IL-5 and eotaxin intrinsically modulate IL-13 production from Th2 cells and that these signaling systems are not necessarily independent effector pathways and may also be integrated to regulate aspects of allergic disease.


The Relationship of Hematoma Growth to Red Blood Cell Distribution Width in Patients with Hypertensive Intracerebral Hemorrhage.
Hypertension is a primary risk factor for intracerebral hemorrhage (ICH) and is thought to be responsible for about 55% of all ICH cases. Thus, the primary goal of the study was to examine whether the status of vascular rheological factors upon admission to the hospital was associated with hypertensive ICH growth and early outcomes. Over a 2-year period, the present study evaluated 60 ICH patients who were admitted within the first 12 hours of symptom onset. Brain computed tomography scans were performed at admission and then 24 hours later as a control. Hematoma growth was classified as an volume increase more than 6.5 ml or > 33%, and good outcome was defined using the modified Rankin Scale (mRS) score (? 2 at 3 months). The mean age of the study population was 65.07 ± 11.659 years, with 34 men and 26 women. The leading vascular risk factor was hypertension (86.7%). There were significant associations between the initial red blood cell distribution width (RDW) and hematoma growth (p=0.038). Therefore, hematoma growth in the first 24 hours after symptom onset was significantly related to a poor clinical outcome at 3 months (p = 0.050). The study identified significant relationships between the initial RDW and poor outcome as well as the initial RDW and hypertensive hematoma growth. Additionally, this study demonstrated that these parameters are easily obtainable and could be used to effectively evaluate outcomes in ICH patients.


A Single-Center Retrospective Evaluation of the Use of Oral Factor Xa Inhibitors in Patients With Cerebral Venous Thrombosis.
Clinical practice guidelines recommend that cerebral venous thrombosis (CVT) be managed with long-term anticoagulant therapy using warfarin or low-molecular-weight heparin (LMWH) for 3 to 12 months. However, oral factor Xa inhibitors may offer preferable alternative treatment options for these patients. The primary objective was to determine the effectiveness and safety of rivaroxaban or apixaban compared with warfarin or enoxaparin as long-term anticoagulation therapy for patients with a new diagnosis of CVT. This was a single-center retrospective review of patients with newly diagnosed CVT who received acute and maintenance anticoagulation treatment. Study groups compared patients who received warfarin, enoxaparin, or an oral factor Xa inhibitor as their maintenance anticoagulant. The primary outcome was recurrent thrombotic events while on anticoagulation. Secondary outcomes included modified Rankin Scale, improved cerebral venous sinus opacification, duration of anticoagulant therapy, bleeding events during anticoagulant therapy, and adverse effects. A total of 119 patients were included in the analysis: warfarin (89), enoxaparin (11), and oral factor Xa inhibitor (19). The risk of recurrent thrombotic events were 11.2%, 0%, and 10.5% in the warfarin, enoxaparin, and oral factor Xa inhibitor treatment groups, respectively (P = 0.7635). There were no significant between-group differences observed regarding any of the secondary outcomes. Although the sample size is limited, these findings indicate that oral factor Xa inhibitors are a reasonable treatment option for patients with CVT. There was a trend toward more persistent symptoms in patients on warfarin, suggesting a potential improvement in recovery among patients that received an oral factor Xa inhibitor.


Pilomatrixoma: a common tumour of head and neck, but rarely reported.
Pilomatrixoma is a benign subcutaneous tumour arising from the sebaceous glands. Mutation in the CTNNB1 gene is seen, suggesting beta-catenin misregulation may be the cause of pilomatrixoma. The preoperative diagnosis may be improved by the awareness of the fact that pilomatrixoma is a common and benign skin tumour of the head and neck region. It presents as a well-defined mass, which may be firm to hard in consistency, usually attached to the skin, but not to the underlying tissue. The colour of overlying skin appears a reddish-brown tinge, indicating that it could be a case of pilomatrixoma. Here, we report a case of pilomatrixoma of the cheek in a woman along with the CT findings and histopathological appearances. Dental surgeons should consider it as one of the differential diagnosis in superficial head and neck swelling with calcification.


Tetrapodal amidoxime ligands I. Coordination isomerism due to self-complementary dimerization of a pyramidal cobalt(III) coordination module.
A bis-μ-amidoximato-bridged cobalt(III) dimer obtained with a new tetrapodal ligand possesses interesting structural parameters as a consequence of intramolecular hydrogen bonding intentionally built into the complex. Its synthesis and properties are described. The new ligand type combines attributes of two previously described ligand classes: It binds a metal ion in a tetrapodal pentadentate fashion and forms a pseudomacrocycle through hydrogen bonds, characteristic of chelating oxime ligands. Coordination isomerism, which is a consequence of dimer formation, has been analyzed by means of X-ray crystallography and carbon-13 nuclear magnetic resonance spectroscopy.


Stress, suicide, and military service during Operation Uphold Democracy.
The initial weeks of Operation Uphold Democracy were marked by a high rate of major axis I disorders among soldiers presenting for treatment at the 528th Combat Stress Center. The mission to Haiti was also marked by three suicides during the first 30 days. The author explores psychodynamic sources of stress that may have contributed to the high level of psychiatric acuity observed during that first month. Then, distinctions are made between psychiatric conditions that present in combat and those that occur on peacekeeping missions.


Comparative study of serine-plasmalogens in human retina and optic nerve: identification of atypical species with odd carbon chains.
The objective of this work was to detect and identify phosphatidylserine plasmalogen species in human ocular neurons represented by the retina and the optic nerve. Plasmalogens (vinyl-ether bearing phospholipids) are commonly found in the forms of phosphatidylcholine and phosphatidylethanolamine in numerous mammalian cell types, including the retina. Although their biological functions are unclear, the alteration of cellular plasmalogen content has been associated with several human disorders such as rhizomelic chondrodysplasia punctata Type 2 and primary open-angle glaucoma. By using liquid chromatography coupled to high-resolution and tandem mass spectrometry, we have identified for the first time several species of phosphatidylserine plasmalogens, including atypical forms having moieties with odd numbers of carbons and unsaturation in sn-2 position. Structural elucidation of the potential phosphatidylserine ether linked species was pursued by performing MS(3) experiments, and three fragments are proposed as marker ions to deduce which fatty acid is linked as ether or ester on the glycerol backbone. Interpretation of the fragmentation patterns based on this scheme enabled the assignment of structures to the m/z values, thereby identifying the phosphatidylserine plasmalogens.


Repression of BLADE-ON-PETIOLE genes by KNOX homeodomain protein BREVIPEDICELLUS is essential for differentiation of secondary xylem in Arabidopsis root.
Repression of boundary genes by KNOTTED1-like homeodomain transcription factor BREVIPEDICELLUS promotes the differentiation of phase II secondary xylem in Arabidopsis roots. Plant growth and development relies on the activity of meristems. Boundaries are domains of restricted growth that separate forming organs and the meristem. Class I KNOX homeodomain transcription factors are important regulators of meristem maintenance. Members of this class including BREVIDICELLUS also called KNOTTED-LIKE FROM ARABIDOPSIS THALIANA1 (BP/KNAT1) fulfill this function in part by spatially regulating boundary genes. The vascular cambium is a lateral meristem that allows for radial expansion of organs during secondary growth. We show here that BP/KNAT1 repression of boundary genes plays a crucial role in root secondary growth. In particular, exclusion of BLADE-ON-PETIOLE1/2 (BOP1/2) and other members of this module from xylem is required for the differentiation of lignified fibers and vessels during the xylem expansion phase of root thickening. These data reveal a previously undiscovered role for boundary genes in the root and shed light on mechanisms controlling wood development in trees.


[Platelet-rich plasma (platelet gel) in secondary alveoloplasty in cleft patients].
The use of Platelet-rich plasma (platelet gel--PRP--) was introduced in the oral and maxilofacial surgery 10 years ago. Its good results are due to the quickly generation of new bone and the acceleration of the period of surgical scar formation. Its employment in the alveolar reconstruction of the cleft patient is not still consistent and the works published in the literature are infrequent. The objective of this preliminary study is presenting our experience with the use of PRP in the reconstruction of the alveolar congenital defects of cleft patients. Between July 2002 and January 2004 were operated 14 patients with congenital alveolar cleft carrying out an standard secondary alveoloplasty. In 12 cases was employed cancellous bone of iliac crest and in two cases the donor area was tibial. The laboratory of Hematology of our Hospital prepared the plasma gel rich in platelets by means of a double centrifuge of autologous serum of the patient. After adding calcium a rich gel in platelets in approximate quantities of 1-2 ml was obtained. In this preliminary report we have studied the aspect of the surgical injury the 3er postoperative day, and the first and second weeks after intervention. These results were compared carried out previously with the Standard alveoloplasty without use of the PRP. Likewise we evaluated the bony density by means of intraoral Rx at 3 and 6 months postoperative. The injury of the alveoloplasty healed more quickly in the patients in which plasma enriched gel was employed. The patients referred less pain and edema in the first days of the postoperative period. The alveolar bony regeneration was faster to the 3 months, though the result was similar in the intraoral Rx to the 6 months. The necessary quantities of cancellous bone were smaller in all cases in which the plasma enriched gel was used (30% less). The use of Platelet-rich plasma (PRP) is a valid protocol for the reconstruction in patients with congenital alveolar clefts. Its low price and morbidity do it recommendable for its employment in cleft patients.


Toward an Asbestos Ban in the United States.
Many developed countries have banned the use of asbestos, but not the United States. There have, however, been multiple efforts in the US to establish strict exposure standards, to limit asbestos use, and to seek compensation through the courts for asbestos-injured workers' In consequence of these efforts, asbestos use has declined dramatically, despite the absence of a legally mandated ban. This manuscript presents a historical review of these efforts.


Blood pressure response to standing is a strong determinant of masked hypertension in young to middle-age individuals.
The pathophysiologic mechanisms of masked hypertension are still debated. The aim of this study was to investigate whether the blood pressure response to standing is a determinant of masked hypertension in young individuals. We studied 1078 individuals (mean age 33.2 ± 8.5 years) with stage-1 untreated hypertension at baseline. Orthostatic response was defined as the difference between six SBP measurements in the orthostatic and supine postures. People with a response more than 6.5 mmHg (upper decile) were defined as hyperreactors. After 3 months of follow-up, 24-h ambulatory BP was measured and the participants were classified as normotensives (N = 120), white-coat hypertensive individuals (N = 168), masked hypertensive individuals (N = 166) and sustained hypertensive individuals (N = 624). In 591 participants, 24-h urinary epinephrine was also measured. Orthostatic response was an independent predictor of masked hypertension after 3 months (P = 0.001). In the whole group, the odds ratio for the Hyperreactors was 2.5 [95% confidence interval (95% CI) 1.5-4.0, P < 0.001]. In the participants stratified by orthostatic response and urinary epinephrine, the odds ratio for masked hypertension was 4.2 (95% CI, 1.8-9.9, P = 0.001) in the hyperreactors with epinephrine above the median and was 2.6 (95% CI, 0.9-7.3, P = 0.069) in those with epinephrine below the median. The association between orthostatic response and masked hypertension was confirmed in the cross-sectional analysis after 3 months (P < 0.001). The present findings indicate that hyperreactivity to standing is a significant determinant of masked hypertension. The odds ratio for masked hypertension was even quadrupled in people with an orthostatic response more than 6.5 mmHg and high urinary epinephrine suggesting a role of sympathoadrenergic activity in the pathogenesis of masked hypertension.


The use of computer imaging technology to facilitate the capture of feedlot necropsy information.
The collection of necropsy information is an integral component of veterinary feedlot consulting. Computer imaging technology can be employed to facilitate the capture of feedlot necropsy data. A digital camera is used to capture necropsy images. Subsequently, the images are electronically transferred to a central site for veterinary interpretation and diagnosis.


Set regulation in asexual and sexual Plasmodium parasites reveals a novel mechanism of stage-specific expression.
Transmission of the malaria parasite depends on specialized gamete precursors (gametocytes) that develop in the bloodstream of a vertebrate host. Gametocyte/gamete differentiation requires controlled patterns of gene expression and regulation not only of stage and gender-specific genes but also of genes associated with DNA replication and mitosis. Once taken up by mosquito, male gametocytes undergo three mitotic cycles within few minutes to produce eight motile gametes. Here we analysed, in two Plasmodium species, the expression of SET, a conserved nuclear protein involved in chromatin dynamics. SET is expressed in both asexual and sexual blood stages but strongly accumulates in male gametocytes. We demonstrated functionally the presence of two distinct promoters upstream of the set open reading frame, the one active in all blood stage parasites while the other active only in gametocytes and in a fraction of schizonts possibly committed to sexual differentiation. In ookinetes both promoters exhibit a basal activity, while in the oocysts the gametocyte-specific promoter is silent and the reporter gene is only transcribed from the constitutive promoter. This transcriptional control, described for the first time in Plasmodium, provides a mechanism by which single-copy genes can be differently modulated during parasite development. In male gametocytes an overexpression of SET might contribute to a prompt entry and execution of S/M phases within mosquito vector.


From the cradle to the grave: activities of GATA-3 throughout T-cell development and differentiation.
GATA family transcription factors play multiple vital roles in hematopoiesis in many cell lineages, and in particular, T cells require GATA-3 for execution of several developmental steps. Transcriptional activation of the Gata3 gene is observed throughout T-cell development and differentiation in a stage-specific fashion. GATA-3 has been described as a master regulator of T-helper 2 (Th2) cell differentiation in mature CD4(+) T cells. During T-cell development in the thymus, its roles in the CD4 versus CD8 lineage choice and at the β-selection checkpoint are the best characterized. In contrast, its importance prior to β-selection has been obscured both by the developmental heterogeneity of double negative (DN) 1 thymocytes and the paucity of early T-lineage progenitors (ETPs), a subpopulation of DN1 cells that contains the most immature thymic progenitors that retain potent T-lineage developmental potential. By examining multiple lines of in vivo evidence procured through the analysis of Gata3 mutant mice, we have recently demonstrated that GATA-3 is additionally required at the earliest stage of thymopoiesis for the development of the ETP population. Here, we review the characterized functions of GATA-3 at each stage of T-cell development and discuss hypothetical molecular pathways that mediate these functions.


Assessing Moral Judgements in Veterinary Students: An Exploratory Mixed-Methods Study from Germany.
Although veterinary ethics is required in veterinary curricula and part of the competencies expected of a trained veterinary surgeon according to the European Association of Establishments for Veterinary Education (EAEVE), knowledge concerning the effects of ethics teaching and tools evaluating moral judgement are scarce. To address this lack of tools with a mixed-methods approach, a questionnaire with three case scenarios presenting typical ethical conflicts of veterinary practice was administered to two groups of veterinary students (one had taken ethics classes, one did not). The questionnaire contained both open-ended and closed questions and was analysed qualitatively and quantitatively. The qualitative part aimed at revealing different argumentation patterns between the two groups, whereas the quantitative part focused on the students' approval of different roles and attitudes possibly relating to veterinarians. The results showed no major differences between both groups. However, answering patterns suggest a clear diversity among the students in their perception of morally relevant factors and the veterinary profession. Awareness of morally challenging elements of their profession was presented by students of both groups. With this exploratory study, the application of an innovative mixed-methods tool for evaluating the moral judgement of veterinary medical students is demonstrated.


Cardiovascular depressant effects of the kappa opioid receptor agonists U50488H and spiradoline mesylate.
The cardiovascular effects of the selective kappa opioid receptor agonists U50488H and spiradoline mesylate were examined in pentobarbital-anesthetized mongrel dogs and chloralose-anesthetized cats. In the dog studies, U50488H (0.01-3.0 mg/kg, i.v.) produced a dose-related depression in mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), left ventricular dp/dt, and heart rate. These effects were completely reversed by a 1 mg/kg i.v. dose of the opioid receptor antagonist naloxone. A second kappa agonist, spiradoline mesylate, also produced a naloxone-reversible cardiovascular depression. Furthermore, the compound did not interfere with the positive inotropic or hypertensive effects of norepinephrine (1 microgram/kg, i.v.), showing that the cardiovascular depressant effects of the kappa agonist are unrelated to interference with alpha- or beta-adrenergic receptor mechanisms. In normal cats anesthetized with chloralose, which produces less depression of sympathetic tone than does pentobarbital, spiradoline mesylate did not decrease the MAP in i.v. doses up to 1.0 mg/kg. However, a dose-related increase in sympathetic nerve discharge (SND) was observed (+290% at 1.0 mg/kg). In contrast, in baroreceptor-denervated cats, spiradoline mesylate caused a dose-related hypotensive effect with no change in SND. These results show that the cardiovascular effects of the kappa agonists are peripherally mediated and that reflex sympathetic activity, if uncompromised, can produce a full compensation.


Route dependent effects of 2-chloroadenosine and theophylline in isolated perfused guinea pig hearts.
The adenosine hypothesis for the metabolic regulation of coronary blood flow remains controversial, in part because of differences in results obtained between intravascular and endogenously released adenosine. The main objective of this study was to compare the responses of coronary perfusate flow to intravascular and pericardial 2-chloroadenosine and theophylline. The periocardial route was used to simulate physiological conditions. An isolated, perfused guinea pig heart preparation was used. Hearts were submerged in 10 ml Krebs-Ringer bicarbonate solution having the same chemical composition as that used to perfuse the coronary vasculature. The submersion fluid was assumed to simulate myocardial interstitial fluid. Hearts were divided into four experimental groups. The first group (n = 6) was used to check that the submerged hearts functioned normally. The second group (n = 12) was used to compare differences between intravascular and pericardial 2-chloroadenosine. The third group (n = 16) was used to evaluate the two routes of administration of 2-chloroadenosine on transmural distribution of coronary flow. The fourth group (n = 24) was used to test the effects of hypoxia in the absence and presence of pericardial theophylline. Hearts were obtained from adult guinea pigs of either sex weighing 300-400 g. All submerged hearts were functionally stable for at least 30 min. Pericardial 2-chloroadenosine was about 1000 times less potent than intravascular 2-chloroadenosine in relaxing the coronary vasculature. Venous concentrations of pericardial 2-chloroadenosine were significantly greater than those of intravascular 2-chloroadenosine, at 0.72(SEM 0.17) v 0.18(0.02) mumol.litre-1. Pericardial 2-chloroadenosine caused more uniform distribution of radiotracer microspheres than intravascular 2-chloroadenosine. Finally, pericardial theophylline significantly attentuated the vasodilator response to hypoxia but not to exogenous adenosine. The isolated submerged heart is a viable preparation for investigating the adenosine hypothesis. Our results suggest that intravascular adenosine does not simulate physiological conditions of interstitial adenosine release and action.


Novel trends in engineered milk products.
Food engineering within the dairy sector is an ever developing field of study purely based on the application of engineering principles and concepts to any aspect of dairy product manufacturing and operations. The last 25 years of science and technology devoted to milk and milk products have led to major advances. The purpose of this paper is to review the history and current status of some engineered milk products and to speculate regarding future trends. Much of the advancement has been directed towards production capacity, mechanisation, automation, hygiene within the processing plant, safety, extensions in shelf life, and new product introductions that bring variety and convenience for the consumer. Significant advancements in product quality have been made, many of these arising from improved knowledge of the functional properties of ingredients and their impact on structure and texture. In addition, further improvements focused on energy efficiency and environmental sustainability have been made and will be needed in the future.


Second moment of the Husimi distribution as a measure of complexity of quantum states.
We propose the second moment of the Husimi distribution as a measure of complexity of quantum states. The inverse of this quantity represents the effective volume in phase space occupied by the Husimi distribution, and has a good correspondence with chaoticity of classical system. Its properties are similar to the classical entropy proposed by Wehrl, but it is much easier to calculate numerically. We calculate this quantity in the quartic oscillator model, and show that it works well as a measure of chaoticity of quantum states.


Hydrodynamics of pair-annihilating disclination lines in nematic liquid crystals.
The pair annihilation of straight line defects with strength +/-1/2 in bulk nematic systems is studied numerically, considering a full coupling of orientational degrees of freedom and hydrodynamics. This work is based on the generalization of the Ericksen-Leslie theory to the tensor order parameter as proposed by Qian and Sheng [T. Qian and P. Sheng, Phys. Rev. E 58, 7475 (1998)]. The approach is particularly suited for the late stages of the annihilation process. It is confirmed that the +1/2 disclination line moves considerably faster than the -1/2 one (e.g., twice as fast) due to the hydrodynamic flow. Symmetries of the important stress tensor terms upon inverting the sign of the winding number and performing a homogeneous in-plane rotation of the Q-tensor eigensystem are discussed. The stress tensor terms that dominantly contribute to the advective flow and to the flow asymmetry are identified.


Association of urinary caffeine and caffeine metabolites with cardiovascular disease risk in adults.
Studies have shown that the consumption of a moderate amount of caffeine is associated with a reduced risk of cardiovascular disease (CVD) and may even be protective against CVD. The purpose of the current study was to evaluate the association between urinary caffeine and its related metabolites and CVD risk in a national representative sample of US adults. We analyzed cross-sectional data from the US National Health and Nutrition Examination Survey from 2009 and 2010. The associations between the levels of urinary caffeine metabolites and self-reported CVD, including congestive heart failure, coronary heart disease, angina, heart attack, and stroke, were examined separately in men and women using multivariate logistic regression models adjusted for covariates. In total, 1916 participants (910 men and 1006 women) were included in the analysis. Among women, the odds ratios of CVD in the highest quartiles of 1,3-dimethylxanthine and 1,3,7-trimethylxanthine were 0.33 (95% confidence interval [CI], 0.12-0.92) and 0.35 (95% CI, 0.13-0.93), respectively, compared with the lowest quartiles. Each one-unit (µmol/L) increase in theophylline concentration was associated with a 0.24-mg/dL increase in high-density lipoprotein cholesterol in the fully adjusted model. Among men, no significant association was observed between urinary caffeine metabolites and CVD. Regarding the subtypes of CVD, compared with women in the lowest quartile for 1,3-dimethylxanthine and 1,3,7-trimethylxanthine, the odds of coronary heart disease decreased by 90% (95% CI, -99% to -11%) and 97% (95% CI, -99% to -47%), respectively, in those in the highest quartile. Urinary 1,3-dimethylxanthine and 1,3,7-trimethylxanthine were significantly and inversely associated with CVDs in women. Additional studies are needed to further confirm the results of this study and explore the underlying mechanisms.<END ABSTRACT>.


Bilateral adrenal primary tumor in Stage 4S neuroblastoma: The Italian experience and review of the literature.
Bilateral adrenal neuroblastoma (NB) is rare and is mainly stage 4S. Its incidence, presenting features, management, and outcome have not been fully defined yet. We searched the Italian NB Registry (RINB) for stage 4S NB infants with bilateral adrenal primary tumor to compare them with stage 4S NB with unilateral tumor. Between 1979 and 2016, the RINB enrolled 3731 NB patients aged 0-18 years including 317 infants (8.5%) diagnosed with stage 4S NB. Eleven/317 (3.5%) had a bilateral adrenal primary tumor (Group 1) and 190/317 (59.9%) had a unilateral tumor (Group 2). Group 1 infants were significantly younger (51 vs. 89 days) but were comparable with Group 2 for any other presenting features. In the absence of specific protocols, upfront treatment was based on symptoms, size of adrenal tumors, and biology, and consisted of observation in 5 cases, radiotherapy in one, chemotherapy in 2, and surgery in 3. Five/11 developed progression and 2 of them, both with MYCN amplification, died. The 5-year EFS rates of Group 1 and 2 were 54.5% vs. 73.3% (P=.14) and 5-year OSs were 81.8% and 89.4%, respectively (P=.44). Our data support the hypothesis that 4S NB infants with bilateral adrenal tumors can have favorable outcome with personalized therapeutic approach. The three patients with MYCN amplified tumor benefited from upfront aggressive chemotherapy, in accordance with current protocols. Because of the rarity of this intriguing form of neuroblastoma, collaborative prospective studies are warranted, especially in view of gaining a better insight on its biological and genetic features.


[Standardized interfaces in gynecologic-obstetric documentation].
Since the introduction of the German GSG-law the usage of software for data recording in clinical departments has substantially grown. These systems should be able to exchange or replicate data, but in most cases the corresponding interfaces are absent or only incompletely implemented. The existence of functional, system-independent interfaces in a CSV-or HL-7-compatible format is a minimal requirement for modern software and should be considered.


Is the depth correction using the geometric mean really necessary in a 99Tcm-DMSA scan in the paediatric population?
Determination of the left to right dimercaptosuccinic acid (DMSA) uptake ratio is theoretically one of the easiest quantitative procedures in nuclear medicine. The quantification can be performed on the posterior view, with or without the lateral view for correction of kidney depth. The geometric mean can also be determined using both the anterior and the posterior views. The aim of this study was to evaluate the occurrence of remarkable differences in the results from quantification of the relative renal function using the geometric mean and those obtained using the posterior counts only. Moreover, we evaluated to what extent the patient age influenced these differences. We reviewed 328 99Tcm-DMSA scans. The difference between the relative renal function obtained using the posterior view and that obtained using the geometric mean was calculated and analysed statistically. For the purpose of evaluating the value of performing the geometric mean calculation in patients of different ages, patients were divided into four age groups (group I, < or =2 years; group II, 3-9 years; group III, 10-18 years; group IV, >18 years). Using the Student's t test, no statistical differences were found in the relative renal function obtained by the two methods (posterior projection and geometric mean) in groups I (t = 0.01, P = 0.992) and II (t = 1.43, P = 0.155), which consisted of patients younger than 10 years (77% of the patients). In groups III and IV statistical differences were found (t = 2.27, P = 0.028 and t = 2.170, P = 0.038), respectively. We conclude that for children under 10 years it is unnecessary to perform depth correction using the geometric mean except in rare cases of major malformations and position anomalies.


[Ovarian tissue banking--primary results].
Lifelong hormone replacement therapy and infertility often follow oncological therapy in young women due to a dramatic reduction of the primordial follicle reserve. Reimplantation and function of frozen ovarian tissue slices were successfully demonstrated in animal models, but the durability is limited. Cryopreservation of a whole ovary could reestablish an almost normal ovarian function. Experiments with porcine ovaries showed histological viability. Cryopreservation and retransplantation of sheep ovaries should demonstrate success by hormonal response and pregnancy.


Detecting perturbations of a radiation field inside a head-sized phantom exposed to therapeutic carbon-ion beams through charged-fragment tracking.
Noninvasive methods to monitor carbon-ion beams in patients are desired to fully exploit the advantages of carbon-ion radiotherapy. Prompt secondary ions produced in nuclear fragmentations of carbon ions are of particular interest for monitoring purposes as they can escape the patient and thus be detected and tracked to measure the radiation field in the irradiated object. This study aims to evaluate the performance of secondary-ion tracking to detect, visualize, and localize an internal air cavity used to mimic inter-fractional changes in the patient anatomy at different depths along the beam axis. In this work, a homogeneous head phantom was irradiated with a realistic carbon-ion treatment plan with a typical prescribed fraction dose of 3 Gy(RBE). Secondary ions were detected by a mini-tracker with an active area of 2 cm2 , based on the Timepix3 semiconductor pixel detector technology. The mini-tracker was placed 120 mm behind the center of the target at an angle of 30 degrees with respect to the beam axis. To assess the performance of the developed method, a 2-mm thick air cavity was inserted in the head phantom at several depths: in front of as well as at the entrance, in the middle, and at the distal end of the target volume. Different reconstruction methods of secondary-ion emission profile were studied using the FLUKA Monte Carlo simulation package. The perturbations in the emission profiles caused by the air cavity were analyzed to detect the presence of the air cavity and localize its position. The perturbations in the radiation field mimicked by the 2-mm thick cavity were found to be significant. A detection significance of at least three standard deviations in terms of spatial distribution of the measured tracks was found for all investigated cavity depths, while the highest significance (six standard deviations) was obtained when the cavity was located upstream of the tumor. For a tracker with an eight-fold sensitive area, the detection significance rose to at least nine standard deviations and up to 17 standard deviations, respectively. The cavity could be detected at all depths and its position measured within 6.5 ± 1.4 mm, which is sufficient for the targeted clinical performance of 10 mm. The presented systematic study concerning the detection and localization of small inter-fractional structure changes in a realistic clinical setting demonstrates that secondary ions carry a large amount of information on the internal structure of the irradiated object and are thus attractive to be further studied for noninvasive monitoring of carbon-ion treatments.


Inducible overexpression of RUNX1b/c in human embryonic stem cells blocks early hematopoiesis from mesoderm.
RUNX1 is absolutely required for definitive hematopoiesis, but the function of RUNX1b/c, two isoforms of human RUNX1, is unclear. We established inducible RUNX1b/c-overexpressing human embryonic stem cell (hESC) lines, in which RUNX1b/c overexpression prevented the emergence of CD34+ cells from early stage, thereby drastically reducing the production of hematopoietic stem/progenitor cells. Simultaneously, the expression of hematopoiesis-related factors was downregulated. However, such blockage effect disappeared from day 6 in hESC/AGM-S3 cell co-cultures, proving that the blockage occurred before the generation of hemogenic endothelial cells. This blockage was partially rescued by RepSox, an inhibitor of the transforming growth factor (TGF)-β signaling pathway, indicating a close relationship between RUNX1b/c and TGF-β pathway. Our results suggest a unique inhibitory function of RUNX1b/c in the development of early hematopoiesis and may aid further understanding of its biological function in normal and diseased models.


Preparation and characterization of anti-tissue factor single-chain variable fragment antibody for cancer diagnosis.
Tissue factor (TF), which serves as the initiator of the extrinsic blood coagulation cascade, has been found to be overexpressed in various solid tumors, especially brain tumors, pancreatic cancer, and gastric cancer. Overexpression of TF is considered to contribute to the high incidence of thrombotic complications and poor prognosis in patients with such cancers. Therefore, detection or targeting of TF may be a promising approach for the diagnosis and treatment of solid tumors that are known to overexpress the protein. Here, we used the recombinant DNA technology to develop an anti-TF single-chain Fv (scFv) of small size and high affinity for its target. The biochemical characteristics of the anti-TF scFv were evaluated using surface plasmon resonance (SPR) sensing and flow cytometry. The data obtained showed that the affinity of the anti-TF scFv was 2.04 × 10(-8) (KD), and that the protein showed significant binding to the cancer cells. Then, Alexa 647-labeled anti-TF scFv and anti-TF IgG were administered to mice bearing chemically induced spontaneous tumors. The maximum tumor to background ratios of anti-TF scFv and anti-TF IgG were obtained 3 and 24 h after the injections, respectively. This study indicates anti-TF scFv may be suitable as an imaging probe for the diagnosis of solid tumors.


Zebra mussels (Dreissena polymorpha) as indicators of freshwater contamination with lindane.
Zebra mussels are common freshwater mollusks in many European lakes and rivers. Their abundance, wide distribution, and filtering activity make them good candidates to evaluate the contamination of freshwaters with environmental contaminants. The purpose of this work was to determine the kinetics of lindane in zebra mussels and compare laboratory results with in situ measurements. Exposure was conducted in small tanks, under controlled experimental conditions. Our results indicated that mussels accumulated lindane with a bioconcentration factor around 10. They generally reached equilibrium within 4 days. Elimination was rapid but biphasic and the terminal elimination half-life was long (> 168 h). Age of the mussels and temperature also affected the kinetics of lindane in mussels. In the Lake of Geneva, zebra mussels were sampled and showed that mussels accumulated it to significant values (up to 900 ng/g fresh weight) depending on the site and period of sampling. The in situ results, together with the laboratory exposures, showed that freshwater mussels could be used to monitor point sources of pollutants such as lindane over short periods of time (< 1 week).


A Filtration System That Greatly Reduces Aluminum in Calcium Gluconate Injection, USP Used to Prepare Parenteral Nutrition Solutions.
The study objective was to reduce aluminum (Al) in Calcium Gluconate Injection, US Pharmacopeia (USP) used in the preparation of parenteral nutrition (PN) solutions. A flow-through filter containing an immobilized chelator that complexes Al from Calcium Gluconate Injection, USP as it flows through the filter was designed, refined by design modifications, and extensively tested. When a small-volume parenteral vial containing 100 mL of Calcium Gluconate Injection, USP is connected on the inlet side of the filter, and the outlet side is connected to an evacuated receiving vial, the filtered solution is drawn into the receiving vial. This constitutes a complete system to remove Al from Calcium Gluconate Injection, USP. The extent of Al removal is flow rate dependent. At a flow rate of 1 mL/min approximately 85% of the Al was removed from calcium gluconate solution. PN solutions have been reported to deliver 15 to 23 mcg/kg/day Al to neonates. Given that Calcium Gluconate Injection, USP provides 85% of the Al in neonatal PN solutions, removal of 85% of the Al from this source was calculated to reduce Al delivered to most neonates to <5 mcg/kg/day. A point-of-use, self-contained, single-use, disposable, Al-complexing filter has been created. It was calculated to reduce Al delivered in PN solutions by 72%, resulting in daily Al delivery below the level that results in Al accumulation associated with central nervous system and bone toxicity to all but the smallest (<1 kg) infants.


Aminoaciduria as a marker of acute renal transplant rejection--a patient study.
Over 12 months, urine samples were systematically collected from 40 children who underwent renal transplantation for the treatment of end-stage renal disease. Sequential determinations of the excretion of individual amino acids relative to that of creatinine were carried out on 15 subjects. Nine of these (including three who sustained episodes of acute rejection) retained a native kidney in-situ, while in six patients (including three who underwent an episode of acute rejection) both native kidneys had been removed. In both subgroups, the amino acid/creatinine ratios of early morning urine samples were higher shortly before clinical manifestations of acute rejection became evident than in patients who, following renal transplantation, had stable kidney function, chronic graft rejection, or acute tubular necrosis, with one exception: a patient with one native kidney in-situ in whom acute tubular necrosis developed immediately after transplantation. The amino acids showing the greatest increase included Thr, Ser, Gly, and Ala. These values fell dramatically immediately prior to the clinical episode of acute rejection, with Thr, Ala, and Phe showing the most consistent changes. These alterations in urinary amino acid excretion occurred several days before changes in urinary protein excretion or the serum concentrations of urea and creatinine, and may have a role to play in the monitoring of renal transplant recipients.


Mitochondrial dysfunction in amyotrophic lateral sclerosis - a valid pharmacological target?
Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disease characterized by the selective death of upper and lower motor neurons which ultimately leads to paralysis and ultimately death. Pathological changes in ALS are closely associated with pronounced and progressive changes in mitochondrial morphology, bioenergetics and calcium homeostasis. Converging evidence suggests that impaired mitochondrial function could be pivotal in the rapid neurodegeneration of this condition. In this review, we provide an update of recent advances in understanding mitochondrial biology in the pathogenesis of ALS and highlight the therapeutic value of pharmacologically targeting mitochondrial biology to slow disease progression.


The contribution of wet labs in the education of ophthalmologists.
The need for pre-training in experimental eye surgery is considered necessary. It is an essential way to assess trainees in ophthalmology based on their instrument and tissue handling and skills. This article aims to underline this necessity and demonstrate the ocular health professionals' opinion on this issue. 74 participants (45 females and 29 males) were included in the study. Ophthalmology residents, ophthalmologists participated in the wet lab session. The evaluation of the contribution of the wet labs were provided by filling a new questionnaire form. In this way, an interactive questionnaire was developed. Regarding trainees' grading of wet labs' significance as a first step for guiding their surgical career, it was positively correlated with their subjective view of labs' utility to both improve their surgical skills (p = 0.001) and maintain pre-existing ones (p < 0.001). We should also note that all of them (100%) stated that wet labs were necessary during residency, especially in repeated sessions, and that they would recommend them to their colleagues. The surgical skills improved significantly after participation in a wet lab, according to participants, who rated the experience as highly educational. Wet labs can reduce the learning curve of difficult surgical techniques, accelerate the rate for trainees to achieve surgical competency, and treat patients safely and effectively.


[Comparative statistical study of global and local functional respiratory data in 2 groups of patients with pulmonary or pleural tuberculosis].
Two groups of patients with unilateral tuberculosis, either pulmonary (36 cases) or pleural (75 cases) were studied. The clinical and functional results were compared statistically. These two groups were similar in terms of age, sex, tobacco consumption, the radiological unilaterality of their disease and the global reduction of respiratory function (assessed by VC, FEV1, TPC and TLCO), which can be considered to be essentially due to the local disease. In both groups, the radiological image (extent of the lesion, shape of the lesion) is the only clinical factor statistically related to the functional values. The two groups differ in terms of two functional parameters which reflect different pathophysiological mechanisms in the affected lung. The group with pulmonary tuberculosis presents an obstructive ventilatory syndrome (reduction of the FEV1/VC ratio and increase in the RV/TPC ratio), which reflects the bronchial involvement in the disease. The group with pleural tuberculosis presents a pulmonary distention (increased RV/TPC ratio), which is related to disturbed transmission of the pleural pressure to ventilated alveolar zones. Secondly, in the group with pulmonary tuberculosis, the ventilation and perfusion deficits are equivalent, while in the pleural tuberculosis group, the ventilatory deficit is predominant. Thus the global reduction in ventilatory function which is the same in the two group is caused by fundamentally distinct pathophysiological mechanisms in the two groups of patients.


Antigen endocytosis and presentation mediated by human membrane IgG1 in the absence of the Ig(alpha)/Ig(beta) dimer.
Membrane immunoglobulin (mIg) M and D heavy chains possess minimal (KVK) cytoplasmic tails and associate with the Ig alpha/Ig beta (CD79) dimer to achieve surface expression and antigen presentation function. In contrast, the cytoplasmic tail of mIgG is extended by 25 residues (gamma ct). We have tested the possibility that mIgG can perform antigen capture and presentation functions independently of the Ig(alpha)/beta dimer. We show that CD4/(gamma)ct chimeras are efficiently endocytosed partially dependent on a tyrosine residue in (gamma)ct. In addition, human mIgG was expressed on the surface of Ig(alpha)/Ig(beta)-negative non-lymphoid cells and mediated antigen capture and endocytosis. Antigen-specific human mIgG targeted antigen to MIIC-type vesicles in the Ig(alpha)/beta negative melanoma Mel JuSo and augmented antigen presentation 1000-fold, identical to the augmentation seen in Ig(alpha)/beta-positive B-cells expressing the same transfected mIgG. Thus, unlike mIgM, mIgG has autonomous antigen capture and presentation capacity, which may have evolved to reduce or eliminate the BCR's dependence on additional accessory molecules.


Enzyme Promiscuity Prediction Using Hierarchy-Informed Multi-Label Classification.
As experimental efforts are costly and time consuming, computational characterization of enzyme capabilities is an attractive alternative. We present and evaluate several machine-learning models to predict which of 983 distinct enzymes, as defined via the Enzyme Commission (EC) numbers, are likely to interact with a given query molecule. Our data consists of enzyme-substrate interactions from the BRENDA database. Some interactions are attributed to natural selection and involve the enzyme's natural substrates. The majority of the interactions however involve non-natural substrates, thus reflecting promiscuous enzymatic activities. We frame this "enzyme promiscuity prediction" problem as a multi-label classification task. We maximally utilize inhibitor and unlabelled data to train prediction models that can take advantage of known hierarchical relationships between enzyme classes. We report that a hierarchical multi-label neural network, EPP-HMCNF, is the best model for solving this problem, outperforming k-nearest neighbours similarity-based and other machine learning models. We show that inhibitor information during training consistently improves predictive power, particularly for EPP-HMCNF. We also show that all promiscuity prediction models perform worse under a realistic data split when compared to a random data split, and when evaluating performance on non-natural substrates compared to natural substrates. We provide Python code for EPP-HMCNF and other models in a repository termed EPP (Enzyme Promiscuity Prediction) at https://github.com/hassounlab/EPP. Supplementary data are available at Bioinformatics online.


Effect of MHC Linked 7-Gene Signature on Delayed Hepatocellular Carcinoma Recurrence.
Dysregulated immune response significantly affects hepatocellular carcinoma's (HCC) prognosis. Human Leukocyte Antigens are key in devising immune responses against HCC. Here, we investigated how HLAs modulate HCC development at the transcriptomic level. RNA-seq data of 576 patients from two independent cohorts was retrieved. The clinicopathological relevance of all HLA genes was investigated using Fisher-Exact, correlation, and Kaplan-Meier and cox regression survival tests. Clustering of ~800 immune-related genes against HLAs was completed using a ward-agglomerative method. Networks were generated using 40 HLA associated unique genes and hub genes were investigated. HLAs including HLA-DMA, HLA-DMB, HLA-DOA and HLA-DRB6 were associated with delayed recurrence in both discovery (204 HCC cases) and validation (372 HCC cases) cohorts. Clustering analyses revealed 40 genes associated with these four HLAs in both cohorts. A set of seven genes (NCF4, TYROBP, LCP2, ZAP70, PTPRC, FYN and WAS) was found co-expressed at gene-gene interaction level in both cohorts. Furthermore, survival analysis revealed seven HLA-linked genes as predictors of delayed recurrence. Multivariate analysis also predicted that mean expression of 7-gene is an independent predictor of delayed recurrence in both cohorts. We conclude that the expression of 7-gene signature may lead to improved patient prognosis. Further studies are required for consideration in clinical practice.


Emotion regulation training to reduce problematic dietary restriction: An experimental analysis.
Evidence suggests that emotion regulation may be a process relevant to problematic dietary restriction. However, emotion regulation has not been evaluated as an intervention target across a range of restriction severity. This study utilized an experimental design to examine whether targeting emotion regulation reduced problematic dietary restriction. Within a self-identified restrictive sample (n = 72), the effects of an emotion regulation condition (i.e., emotion regulation training) were compared to those of a control condition (i.e., nutrition information training) on dietary restriction indices (i.e., effort to reduce intake on a progressive ratio task, work towards an alternate reinforcer on a progressive ratio task, intake by dietary recall) following a stressor. Exploratory analyses of potential moderators (i.e., restraint, BMI, binge eating and purging status, emotion regulation difficulties) were conducted to examine whether these factors affected the impact of training on dietary restriction. No significant main effects of condition were detected on any outcome measure. However, results were moderated by BMI status. Participants with lower BMIs exerted less effort towards dietary restriction following the emotion regulation condition versus the control condition (p = 0.02). Results suggest that targeting emotion regulation may help to reduce problematic dietary restriction among lower weight individuals.


The strong correlation between ADAM33 expression and airway inflammation in chronic obstructive pulmonary disease and candidate for biomarker and treatment of COPD.
Airway inflammation in patients with chronic obstructive pulmonary disease (COPD) is an amplified response of the normal immune system that occurs as a result of chronic irritation by toxic substances, such as cigarette smoke. This leads to the characteristic pathological changes in the inflammatory cells of COPD patients. ADAM33 has been reported to be involved in the pathogenesis of COPD in East Asia by affecting airway inflammation and other immune responses. The aim of this study was to determine the potential role of ADAM33 (mRNA and soluble levels) as a biomarker of inflammation in COPD patients. This is a case control study using consecutive sampling. The COPD case and control (non-COPD) groups comprised 37 and 29 patients, respectively. We used univariate analysis to assess differences in the parameters between the groups and bivariate analysis to non-parametrically compare these parameters between the two groups. We observed significantly higher mRNA levels of ADAM33 in the COPD patients (10.39 ± 1.76) as compared to that in the non-COPD individuals (6.93 ± 0.39; P < 0.001). The levels of soluble ADAM33 were also significantly higher in the COPD patients (2.188 ± 1.142 ng/ml) compared to the non-COPD individuals (0.487 ± 0.105 ng/ml; P < 0.001). The mRNA and soluble ADAM33 levels were significantly higher in COPD patients compared to those in the parameter-matched non-COPD individuals. Thus, ADAM33 is a potential biomarker and treatment for inflammation in COPD patients.


On the identification of beneficial and detrimental molecular forms of fibrinogen.
Fibrinogen is a central protein in blood coagulation. A functioning circulation system requires a precise balance between fibrin formation and removal, i.e. between the interaction of fibrin(ogen) with thrombogenic and fibrinolytic components of the blood. Fibrinogen and fibrin have also significant roles in wound healing, in tumor growth and metastasis as well as in defense mechanisms. All functions and interactions are mediated by specific structural elements of the molecule. Already in healthy individuals fibrinogen occurs in over a million nonidentical forms due to posttranslational modifications and genetic polymorphism. The various forms may show considerable differences in their functional properties. Alterations in distributions among preexisting forms as well as additional forms have been observed to accompany many types of disease. Furthermore, certain forms have been correlated with an increased risk to acquire disease. Monitoring the levels of various molecular forms is expected to be of considerable diagnostic and prognostic value in many types of disease.


Differential effects of desmoglein 1 and desmoglein 3 on desmosome formation.
The desmoglein plays an important part in the formation of desmosomes. We constructed recombinant adenoviruses containing desmoglein 1 and desmoglein 3 derivatives partly lacking the extracellular domain (desmoglein 1DeltaEC and desmoglein 3DeltaEC, respectively), and full-length desmoglein 1 and desmoglein 3 and studied the involvement of desmoglein 1 and desmoglein 3 in desmosome formation. During low-level expression of desmoglein 3DeltaEC in transduced HaCaT cells, keratin insertion at cell-cell contact sites was only partially inhibited and desmoplakin was partially stained at cell-cell contact sites. Low-level expression of desmoglein 1DeltaEC, however, resulted in complete inhibition of keratin insertion at the cell-cell contact sites, and desmoplakin was stained in perinuclear dots. These results indicate the dominant-negative effect of desmoglein 1DeltaEC on desmosome formation was stronger than that of desmoglein 3DeltaEC. Desmoglein 1DeltaEC coprecipitated plakoglobin to approximately the same extent as desmoglein 3DeltaEC. Therefore, we conclude that the dominant-negative effect of desmoglein 1DeltaEC is not simply due to plakoglobin sequestration. On the other hand, during low-level expression of full-length desmoglein 3 and desmoglein 1, they both colocalized with desmoplakin. During high-level expression, however, keratin insertion at cell-cell contact sites was inhibited in desmoglein 1 but not in desmoglein 3, and desmoplakin was stained at cell-cell contact sites in desmoglein 3 but not in desmoglein 1. These data suggest desmoglein 1 and desmoglein 3 expressed at low level were incorporated into desmosome but at high-level expression, desmoglein 1 disrupted desmosomes but desmoglein 3 did not. Our findings provide biologic evidence that desmoglein 1 and desmoglein 3 play a different functional role in cell-cell adhesion of keratinocytes.


Internal lumbar disc derangement with instability catch from monosegmental discopathy. The forgotten mechanical and kinetic surgical back pain syndrome.
This is a retrospective cohort experience reported with concurrent survey PROM outcomes. To describe the results of open PLIF reconstruction for a select group of mechanical back pain patients who have mono- or bi-segmental discopathy on MRI imaging, a clinical history of repeated severe and disabling acute mechanical back pain symptoms, and the irregular lumbar motion pattern in returning erect from the flexed position known as the "instability catch". The literature of fusion surgery for back pain relief in "mechanical" back pain reveals inconsistent results and in the majority presents only a vague description of these syndromes. Internal Lumbar Disc Degeneration with Instability catch "ILDDIC" may be one subset of these patients who are uniquely benefitted from spine stabilization. The senior author (DAB) in midsummer 2015 began to offer smaller fusion procedures to selected patients on an overnight-stay basis using a standard perioperative care protocol. For practice audit, in December 2020 a mailed survey questionnaire requesting VAS pain scores and SF-36 physical function scores was sent out to all 111 patients who had been treated this way, which group included 30 cases of ILDDIC. We report here on the success of open PLIF reconstruction in achieving back pain relief for these patients. Some 24 of 30 patients returned the mailed survey questionnaire, and the remaining six could not be reached. All 24 responders reported significant relief of back pain and improved physical function, at a mean of 30 months from surgery. Review of the available clinical records (LFU < 1 year) from the six nonresponders also recorded positive early benefit. The diagnosis of ILDDIC requires both imaging and clinical correlates and may define a subgroup of the mechanical back pain population uniquely suited to achieve pain relief through lumbar fusion.


The doublesex proteins of Drosophila melanogaster bind directly to a sex-specific yolk protein gene enhancer.
The doublesex (dsx) gene of Drosophila melanogaster encodes both male-specific and female-specific polypeptides, whose synthesis is regulated by alternative sex-specific splicing of the primary dsx transcript. The alternative splicing of the dsx mRNA is the last known step in a cascade of regulatory gene interactions that involves both transcriptional and post-transcriptional mechanisms. Genetic studies have shown that the products of the dsx locus are required for correct somatic sexual differentiation of both sexes, and have suggested that each dsx product functions by repressing expression of terminal differentiation genes specific to the opposite sex. However, these studies have not shown whether the dsx gene products function directly to regulate the expression of target genes, or indirectly through another regulatory gene. We report here that the male- and female-specific DSX proteins, expressed in E.coli, bind directly and specifically in vitro to three DNA sequences located in an enhancer region that regulates female-specific expression of two target genes, the yolk protein genes 1 and 2. This result suggests strongly that dsx is a final regulatory gene in the hierarchy of regulatory genes controlling somatic sexual differentiation.


Receptor-mediated endocytosis of carcinoembryonic antigen by rat liver Kupffer cells.
In vivo, carcinoembryonic antigen (CEA) is removed from the circulation by the liver Kupffer cells. Immunologically identifiable CEA is transferred from these macrophages to the hepatocytes, where degradation is completed. Circulatory clearance of CEA is specific, rapid [t1/2 = 3.7 +/- 0.9 (S.D.) min], and saturable. In vitro, Kupffer cells take up CEA by a saturable process which is time/temperature dependent and colchicine sensitive. Isolated Kupffer cells endocytose CEA with an apparent Km of 6 X 10(-8) M. There are approximately 16,000 CEA binding sites per cell. Nonspecific cross-reacting antigen (NCA), a glycoprotein structurally similar to CEA, is recognized with lower affinity by the same receptor. Endocytosis is independent of the nonreducing terminal sugars on the molecule: CEA modified by Smith degradation inhibits Kupffer cell recognition of native CEA. Since performic acid oxidized CEA also inhibits endocytosis, receptor binding is similarly independent of intact protein conformation. Isolated Kupffer cells have mannose and/or N-acetyl glucosamine receptor activity but do not internalize CEA by that mechanism. Galactose-terminated glycoproteins impede CEA and NCA clearance in vivo but not Kupffer cell endocytosis in vitro. Radiolabeled CEA released from isolated Kupffer cells following endocytosis shows no apparent molecular weight change. However, the released CEA contains species with higher isoelectric points, suggesting that perhaps the removal of sialic acid and the resulting exposure of galactose residues mediate the subsequent transfer to the hepatocyte.


Effects of cholinergic enhancement on conditioning-related responses in human auditory cortex.
It has previously been shown that cholinergic blockade attenuates conditioning-related neuronal responses in human auditory cortex. The present study was conducted to investigate the effect of cholinergic enhancement on such experience-dependent cortical responses. The cholinesterase inhibitor physostigmine, or a placebo control, were continuously infused into healthy young volunteers, during differential aversive conditioning whilst brain activity was measured using event-related functional magnetic resonance imaging (fMRI). Volunteers were presented with two tones, one of which (CS+) was conditioned by pairing with an electrical shock whereas the other was always presented without the shock (CS-). Conditioning-related activations, expressed as an enhanced blood oxygenation level dependent (BOLD) response to the salient CS+, were evident in left auditory cortex under placebo but not under physostigmine. This absence of conditioning-related activations under physostigmine was due to enhanced responses to the CS- under physostigmine as compared to placebo. We suggest that an overactive cholinergic system leads to increased processing of behaviourally irrelevant stimuli and thus attenuates differential conditioning-related cortical activations.


Solid-State Na Metal Batteries with Superior Cycling Stability Enabled by Ferroelectric Enhanced Na/Na3 Zr2 Si2 PO12 Interface.
Solid-state metal batteries are attracting unprecedented concern because of their high energy density and safety. However, their service life, especially at high specific density, is hindered by the undesirable reversibility of metal anodes, owing to the inhomogeneous ion distribution and awkward charge transfer dynamics at the electrode/electrolyte interface. In this work, it is well demonstrated that ferroelectric phase BaTiO3 reinforced Na3 Zr2 Si2 PO12 ceramic electrolyte can deconcentrate the distribution of charge transfer and self-accelerate Na+ migration at the Na/Na3 Zr2 Si2 PO12 interface upon cycling, realizing a compact Na deposition morphology together with a high critical current density (1.05 mA cm-2 at ambient conditions). Assembled symmetric cells based on the proposed composite electrolyte render stable cycling up to 1000 h at 0.3 mA cm-2 . Specifically, the all solid-state sodium metal batteries paired with Na3 V1.5 Cr0.5 (PO4 )3 cathode material can deliver a capacity of 95 mAh g-1 at 100 mA g-1 and maintain 84.4% of the initial capacity after 400 cycles. This excellent electrochemical performance clearly confirm the feasibility of the introduction of ferroelectric BaTiO3 to suppress the dendrite nucleation and Na propagation within ceramic electrolyte. This research offers new insight into the rational design of inorganic electrolyte, revealing dendrite-free and long-term all-solid-state sodium batteries.


A PCR method based on 18S rRNA gene for detection of malaria parasite in Balochistan.
To establish a polymerase chain reaction method based on 18S ribosomal ribonucleic acid gene for the detection of plasmodium deoxyribonucleic acid in patients suffering from malaria symptoms. This cross-sectional study was conducted from September 2013 to October 2014 in district Quetta of Pakistan's Balochistan province. Blood samples were collected from patients suffering from general symptoms of malaria. A polymerase chain reaction-based technique was applied for the diagnosis of malaria and detection of responsible species in the patients who were suspected to carry the parasite. Performance of this polymerase chain reaction method was compared against the microscopy results. Parasite number was also calculated for microscopy positive samples.All samples after the genomic deoxyribonucleic acid isolation were subjected to polymerase chain reaction amplification and agarose gel electrophoresis. Of the 200 samples, 114(57%) were confirmed as positive and 86(43%) as negative for malaria by microscopy. Polymerase chain reaction identified 124(62%) samples as positive and 76(38%) as negative for malaria. The comparative analysis of both diagnostic methods confirmed 109(54.5%) samples as positive by both techniques. Besides, 5(6.58%) samples were identified as false positive and 15(12.1%) samples as false negative by polymerase chain reaction. Sensitivity, specificity and positive predictive values for polymerase chain reaction in comparison to microscopy were 87.98%, 93.42% and 96%, respectively. Polymerase chain reaction-based methods in malaria diagnosis and species identification were found to be more effective than other techniques.


The vaccine-site microenvironment: impacts of antigen, adjuvant, and same-site vaccination on antigen presentation and immune signaling.
A goal of cancer vaccines is to induce strong T cell responses to tumor antigens, but the delivery method, schedule, and formulation of cancer vaccines have not yet been optimized. Adjuvants serve to increase the immune response against vaccine antigens. However, little is known about the impact of adjuvants plus antigen and their delivery schedule on the immunologic milieu in the vaccine-site microenvironment (VSME). We hypothesized that antigen processing and presentation may occur directly in the VSME, that adding the toll-like receptor 3 (TLR3) agonist polyICLC (pICLC) would enhance markers of immune activation, and that the immune signatures would be enhanced further by repeated vaccination in the same skin site rather than after multiple vaccines in different skin locations. Using RNA sequencing, we evaluated VSME biopsies from patients undergoing subcutaneous/intradermal peptide vaccination against melanoma, with incomplete Freund's adjuvant (IFA) with or without pICLC. Differential gene expression analyses and gene set enrichment analyses were performed using R. False discovery rate corrected p values <0.05 were considered significant. We found that addition of peptide antigens to IFA enhanced antigen presentation pathways and a tertiary lymphoid structure gene-signature locally at the VSME. Addition of pICLC to IFA + peptide induced an immunologically favorable VSME 1 week after injection but had little impact on the VSME after three injections, compared with IFA + peptide alone. Repeated same-site injection of IFA + peptide antigens induced a VSME with more dendritic cell activation, Th1 dominance, and TLR adaptor protein gene expression than that induced by injections at different, rotating skin locations. These data suggest that the vaccine-site itself may be a critically important location contributing to vaccine immunity rather than just the draining lymph node, that IFA induces a favorable VSME with TLR agonist being most beneficial early in the vaccine course, and that same-site injections lead to persistent stimulation of immune pathways that may be beneficial in eliciting antigen specific T cell expansion.


[A patient with systemic lupus erythematosus and lupus nephritis: a 12-year follow-up].
Systemic lupus erythematosus (SLE) is a chronic immunological disease causing a significant morbidity and mortality in younger women and involving several organs and systems, most often the kidneys, being consequently the incidence of lupus nephritis (LN) about 60%. We reported a 57 year-old patient with the diagnosed SLE in 1995. Pathohistological analysis of kidney biopsy revealed LN type V. The patient was treated with corticosteroid pulses and azathioprine during one year. A remission was achieved and maintained with prednisone, 15 mg daily. Nephrotic relapse was diagnosed in 2006 and the second kidney biopsy revealed recent kidney infarction due to extensive vasculitis. Soon, a cerebrovascul insult developed and CT-scan revealed endocranial infarctus. The patient was treated with corticosteroids and cyclophosphamide pulses (totally VI monthly pulses), and also with low-molecular heparine, anticoagulants and salicylates because of the right leg phlebothrombosis. After the pulses, the patient was adviced to take prednisone 20 mg daily and azothioprine 100 mg daily, and 6 months later mycophenolate mofetil because of persistent active serological immunological findings (ANA 1:320) and nephrotic syndrome. Mycophenolate mofetil was efficient in inducing and maintaining remission of nephrotic syndrome. The aim of LN treatment is to achieve and maintain remission, improve patients' outcome, reduce the toxicity of immunosuppressive drugs and the incidence of relapses.Mycophenolate mofetil was shown to be efficient in inducing and maintaining remission of nephrotic syndrome in the frame of LN.