N-Acetyl-D-Glucosamine Kinase Promotes the Axonal Growth of Developing Neurons.
N-acetyl-D-glucosamine kinase (NAGK) plays an enzyme activity-independent, non-canonical role in the dendritogenesis of hippocampal neurons in culture. In this study, we investigated its role in axonal development. We found NAGK was distributed throughout neurons until developmental stage 3 (axonal outgrowth), and that its axonal expression remarkably decreased during stage 4 (dendritic outgrowth) and became negligible in stage 5 (mature). Immunocytochemistry (ICC) showed colocalization of NAGK with tubulin in hippocampal neurons and with Golgi in somata, dendrites, and nascent axons. A proximity ligation assay (PLA) for NAGK and Golgi marker protein followed by ICC for tubulin or dynein light chain roadblock type 1 (DYNLRB1) in stage 3 neurons showed NAGK-Golgi complex colocalized with DYNLRB1 at the tips of microtubule (MT) fibers in axonal growth cones and in somatodendritic areas. PLAs for NAGK-dynein combined with tubulin or Golgi ICC showed similar signal patterns, indicating a three way interaction between NAGK, dynein, and Golgi in growing axons. In addition, overexpression of the NAGK gene and of kinase mutant NAGK genes increased axonal lengths, and knockdown of NAGK by small hairpin (sh) RNA reduced axonal lengths; suggesting a structural role for NAGK in axonal growth. Finally, transfection of 'DYNLRB1 (74-96)', a small peptide derived from DYNLRB1's C-terminal, which binds with NAGK, resulted in neurons with shorter axons in culture. The authors suggest a NAGK-dynein-Golgi tripartite interaction in growing axons is instrumental during early axonal development.


Anxiety, cognitive performance, and cognitive decline in normal aging.
A sample of 704 cognitively intact individuals (M age = 63.7 years) performed a battery of cognitive tests on as many as three occasions, at approximately 3-year intervals. The authors used random effects models to analyze cross-sectional relationships between cognitive performance and state anxiety and longitudinal relationships between cognitive change and neuroticism, after controlling for gender, age, and education. Cross-sectionally, higher state anxiety was associated with poorer performance on Wechsler Adult Intelligence Scale Synonyms, WIT III Analogies, Koh's Block Design, two measures of visual learning (Names and Faces and Thurstone's Picture Memory), and, for men, CVB-Scales Digit Span Test and Card Rotations. In longitudinal models, the main effects for neuroticism were significant for Block Design, Symbol Digit, and Names and Faces, but there were no significant interactions among neuroticism, gender, and time. These results provide some support for Eysenck's processing efficiency theory but none for neuroticism as a risk factor for cognitive decline in normal aging.


In-vitro endothelialization of patches of bovine jugular vein stabilized by dye-mediated photooxidation.
The study aim was to evaluate the cytotoxicity of photooxidatively stabilized bovine jugular vein and the in-vitro endothelialization properties of such material. Bovine jugular veins were cross-linked by dye-mediated photooxidation and cut into 4 cm2 pieces. Ovine jugular vein endothelial cells were isolated, cultured, and then seeded onto the inner surface of bovine jugular vein patches at a density of 1.5 x 10(5) per cm2. Specimens were cultured for five days and then examined using scanning electron microscopy (SEM) and immunohistochemical staining for von Willebrand factor. SEM showed confluent lining of cultured ovine jugular vein endothelial cells similar to native endothelium. Von Willebrand factor staining for cells on the surface of patches was positive. The photooxidatively stabilized bovine jugular vein patches were not cytotoxic, and their in-vitro endothelialization was possible.


Multidentate Ligand-Passivated CsPbI3 Perovskite Nanocrystals for Stable and Efficient Red-Light-Emitting Diodes.
CsPbI3 nanocrystals (NCs) have become a research hot spot in the field of light-emitting diodes (LEDs). Whereas, the long chain ligands with weak affinity to CsPbI3 NCs have prevented their further development and commercialization. Herein, a novel multidentate short ligand tetramethylthiuram disulfide (TMTD) was employed via a ligand exchange process to enhance hole mobility and decrease trap density of the CsPbI3 NCs film. Therefore, TMTD passivated CsPbI3 NCs LED exhibited 20.65% maximum external quantum efficiency and 3861 cd/m2 maximum luminance. Furthermore, TMTD passivated CsPbI3 NCs LED exhibited good operational stability with a 128 min half-lifetime. This strategy using multidentate short ligand passivation provides an effective way to promote perovskite LED development and commercialization.


Structural determinants for vitamin D receptor response to endocrine and xenobiotic signals.
The vitamin D receptor (VDR), initially identified as a nuclear receptor for 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3], regulates calcium metabolism, cellular proliferation and differentiation, immune responses, and other physiological processes. Recently, secondary bile acids such as lithocholic acid (LCA) were identified as endogenous VDR agonists. To identify structural determinants required for VDR activation by 1alpha,25(OH)2D3 and LCA, we generated VDR mutants predicted to modulate ligand response based on sequence homology to pregnane X receptor, another bile acid-responsive nuclear receptor. In both vitamin D response element activation and mammalian two-hybrid assays, we found that VDR-S278V is activated by 1alpha,25(OH)2D3 but not by LCA, whereas VDR-S237M can respond to LCA but not to 1alpha,25(OH)2D3. Competitive ligand binding analysis reveals that LCA, but not 1alpha,25(OH)2D3, effectively binds to VDR-S237M and both 1alpha,25(OH)2D3 and LCA bind to VDR-S278V. We propose a docking model for LCA binding to VDR that is supported by mutagenesis data. Comparative analysis of the VDR-LCA and VDR-1alpha,25(OH)2D3 structure-activity relationships should be useful in the development of bile acid-derived synthetic VDR ligands that selectively target VDR function in cancer and immune disorders without inducing adverse hypercalcemic effects.


Ocular morphology and function in juvenile neuronal ceroid lipofuscinosis (CLN3) in the first decade of life.
CLN3 is a rare lysosomal storage disorder. The majority of the patients suffer from neurological degeneration in the first decade of life leading to death in the second or third decade. One of the first symptoms is a rapid visual decline from retinal degeneration. The aim of this study was to correlate the retinal changes in CLN3 as seen with spectral domain optical coherence tomography (SD-OCT) with functional data in patients in the first years after the subjective onset of ocular symptoms. Three unrelated children aged from 5.6 to 8.8 years, and with molecularly confirmed CLN3, underwent a comprehensive ophthalmological examination including visual acuity, fundus photography, fundus autofluorescence (FAF), electrophysiology (multifocal ERG), Goldmann visual fields, and SD-OCT. A predominant loss of the first and second neuron retinal layers progressing from the macula to the periphery was identifed. The retinal nerve fibre layer (RNFL) displayed gliosis and an irregular lining of the inner limiting membrane. Compared to the preferential reduction of photoreceptor layer thickness in other maculopathies with pan-retinal involvement, the thickness of the first and second neuron layers was reduced simultaneously in CLN3. Functional testing by multifocal ERG reflected the degenerative progress. Semiquantitative evaluation revealed a generally reduced FAF. This is the first detailed morphological evaluation of CLN3 patients in the first years after the subjective onset of ocular symptoms. CLN3 is characterized by an early degeneration predominant of the first and second neuron compared to other macular and generalized retinal dystrophies. Imaging is instrumental for early diagnosis and gene-directed molecular analysis of this fatal disorder.


Effects of apolipoprotein E on the human immunodeficiency virus protein Tat in neuronal cultures and synaptosomes.
Human immunodeficiency virus type 1 (HIV-1)-associated dementia is observed in 20-30% of patients with acquired immunodeficiency syndrome (AIDS). The epsilon4 allele of the apolipoprotein E (APOE) gene currently is thought to play a role as a risk factor for the development of HIV dementia. The HIV protein Tat is neurotoxic and binds to the same receptor as apoE, the low-density lipoprotein receptor-related protein (LRP). In this study, we investigated the role apoE plays in Tat toxicity. Synaptosomes from wild-type mice treated with Tat had increased reactive oxygen species (ROS), increased lipid and protein oxidation, and decreased mitochondrial membrane potential. Synaptosomes from APOE-knockout mice also had increased ROS, increased protein oxidation, and decreased mitochondrial membrane potential, but to a significantly lesser degree. Treatment of synaptosomes with heparinase and Tat increased Tat-induced oxidative stress, consistent with the notion of Tat requiring interaction with neuronal membranes to induce oxidative damage. Human lipidated apoE3 greatly protected neurons from Tat-induced toxicity, whereas human lipidated apoE4 showed no protection. We demonstrated that human apoE3 has antioxidant properties against Tat-induced toxicity. Taken together, the data suggest that murine apoE and human apoE4 act similarly and do not protect the cell from Tat-induced toxicity. This would allow excess Tat to remain outside the cell and interact with synaptosomal membranes, leading to oxidative stress and neurotoxicity, which could contribute to dementia associated with HIV. We show that the antioxidant properties of apoE3 greatly outweigh the competition for clearance in deterring Tat-induced oxidative stress.


Statistical controversies in clinical research: long-term follow-up of clinical trials in cancer.
Long-term follow-up is an important unmet need for the full analysis of new treatments for cancer. Earlier detection of cancer and more effective treatment have led to many more patients surviving for more than 5 and even 10 years, so that evaluating late recurrences and side-effects is an increasingly important issue. This is particularly relevant for oestrogen receptor-positive breast cancer, where the existence of late recurrences is well documented. However, survival for other cancers, notably prostate, colorectal and cervix cancer, has dramatically increased in recent years due to screening and better treatment of early lesions. Trials of preventive therapies have an even greater need for long follow-up. Here, we review these issues and suggest ways in which provision for long-term follow-up can be improved.


Assessment of Immunization to Hepatitis B Vaccine among Children under Five Years in Rural Areas of Taiz, Yemen.
Background. Hepatitis B virus (HBV) infection poses a major health problem worldwide. approximately 1 million deaths annually due to cirrhosis and hepatocellular carcinoma. Objectives. This study was conducted to determine the coverage rate of HBV vaccine and assess the vaccine protective response among children under five years old in rural areas of Yemen. Methods. A cross-sectional study was conducted from January to December 2015 in four districts of countryside Yemen. The target population was children aged from 6 to 59 months. 227 children were enrolled in the study. Questionnaire was used to collect of data. Serum samples were tested for anti-HBs antibodies by enzyme linked immunosorbent assay (ELISA). Anti-HBs level ≥ 10 IU/L was considered a protective response to the vaccine. Results. The coverage rate of HBV vaccine among children was 87.3%. A total of 143 (72.2%) children responded to the vaccine with anti-HBs level ≥ 10 IU/L, while 55 (27.8%) of the children had nonprotective anti-HBs levels of <10 IU/L (P = 0.003). Conclusion. This study revealed a good coverage rate of HBV vaccine in rural areas but the protective rate against HBV infection was moderate. A considerable proportion of vaccinated children should be considered for either revaccination or booster doses.


Inflammatory cytokines and cardiovascular disease.
The designation of atherosclerosis as a chronic inflammatory process represents an interesting paradigmatic shift for cardiologists. The plasma concentrations of interleukin-6 and its hepatic byproduct, C-reactive protein, may reflect the intensity of occult plaque inflammation and the vulnerability to rupture. Monocyte chemoattractant protein-1 and interleukin-8 play a crucial role in initiating atherosclerosis by recruiting monocytes/macrophages to the vessel wall, which promotes atherosclerotic lesions and plaque vulnerability. In addition, circulating levels of these proinflammatory cytokines increase in patients with acute myocardial infarction and unstable angina, but not in those with stable angina. Also, the plasma concentrations of these cytokines increase after percutaneous coronary intervention, causing late restenosis after the procedure. Angiotensin II and other atherogenic factors induce these cytokines in the cardiovascular tissues through the activation of transcription factors, such as nuclear factor-kappaB or peroxisome proliferator-activated receptors. Conversely, HMG-CoA reductase inhibitors (statins) can potently inhibit these proinflammatory factors in the vessels. A small GTP-binding protein, Rho, may be a key molecule to explain the anti-inflammatory effects of statins. Interleukin-10 also exerts anti-inflammatory effects on the cardiovascular tissues, possibly by deactivating proinflammatory cytokines and inducible nitric oxide synthase. Gene therapy using interleukin-10 may be a promising means for untreatable or complicated cases of cardiovascular diseases. Thus, therapeutic modulations of these inflammatory cytokines may be useful in the prevention of atherosclerosis and future cardiovascular events.


Ovine chorionic somatomammotropin and foetal growth.
Ovine chorionic somatomammotropin (oCS) enhances the weight and bone growth of hypophysectomized rats. It acts as a bifunctional hormone, since it binds both to lactogenic and somatotropic receptors. Ovine foetus weight gain is closely related to oCS and oGH serum levels. oCS is able to stimulate somatomedins by foetal liver. Moreover, oCS specific receptors are present in some foetal tissues. So, all these facts involve oCS in foetal growth, whereas pituitary growth hormone intervenes in postnatal growth. A study of structure function relationships between growth hormones and placental hormones is exposed in order to localize somatotropic sites.


Bevacizumab reduces tumor targeting of antiepidermal growth factor and anti-insulin-like growth factor 1 receptor antibodies.
Bevacizumab (antivascular endothelial growth factor [anti-VEGF]) and cetuximab (antiepidermal growth factor receptor [anti-EGFR]) are approved antibodies for treatment of cancer. However, in advanced colorectal cancer, the combination fails to improve survival. As the reason for the lack of activity is unknown, our study aims to determine the effect of bevacizumab on targeting of anti-EGFR and insulin-like growth factor 1 receptor (IGF-1R) antibodies in tumors with single-photon emission computed tomography (SPECT)/CT imaging. Mice with subcutaneous EGFR and IGF-1R-expressing SUM149 xenografts received a single dose of bevacizumab (10 mg/kg) or saline. After 4 days, mice were injected with radiolabeled cetuximab or R1507, an anti-IGF-1R antibody. A control group received a radiolabeled irrelevant IgG (hLL2). Three days later, SPECT/CT images were acquired and mice were dissected to determine the concentration of antibodies in the tissues. Tumors were analyzed immunohistochemically to determine vascular density (CD34), VEGF, EGFR and IGF-1R expression. SPECT/CT imaging revealed that bevacizumab treatment significantly reduced tumor targeting of radiolabeled cetuximab by 40% from 33.1 ± 1.1 %ID/g to 19.8 ± 5.7 %ID/g (p = 0.009) for untreated and bevacizumab-treated tumors, respectively. A similar effect was found for (111) In-R1507: tumor targeting of R1507 decreased by 35%. No significant differences in tumor uptake were observed in mice that received an irrelevant IgG. Uptake in normal organs was not altered by bevacizumab. Immunohistochemical analysis showed that vascular density decreased with 43%, whereas EGFR and IGF-1R expression was unaltered. In conclusion, bevacizumab treatment significantly reduces tumor targeting of anti-EGFR and anti-IGF-1R antibodies. This emphasizes the importance of timing and sequencing of bevacizumab in combination with other antibodies.


Quantitative Proteomics Analysis Revealed Compromised Chicken Dendritic Cells Function at Early Stage of Very Virulent Infectious Bursal Disease Virus Infection.
Chicken dendritic cells (DCs) have been demonstrated to be susceptible to infectious bursal disease virus (IBDV), a causative agent of acute and immunosuppressed disease in young chicks known as infectious bursal disease. Further functional characterization of IBDV-infected DCs of chickens is required to provide a better understanding on the influence of the virus on chicken bone marrow-derived dendritic cells (BM-DCs) following very virulent (vv) IBDV infection. Membrane proteins of BM-DCs were extracted and the proteins were further denatured and reduced before performing labeling with isobaric tags for relative and absolute quantitation. The differential expression protein profiles were identified and quantified using liquid chromatography coupled with tandem mass spectrometry, and later validated using flow cytometry and real-time reverse transcriptase PCR. The analysis has identified 134 differentially regulated proteins from a total of 283 proteins (cutoff values of ≤0.67, ≥1.5, and ProtScore >1.3 at 95% confidence interval), which produced high-yield membrane fractions. The entry of vvIBDV into the plasma membrane of BM-DCs was observed at 3 hr postinfection by the disruption of several important protein molecule functions, namely apoptosis, RNA/DNA/protein synthesis, and transport and cellular organization, without the activation of proteins associated with signaling. At the later stage of infection, vvIBDV induced expression of several proteins, namely CD200 receptor 1-A, integrin alpha-5, HSP-90, cathepsin, lysosomal-associated membrane protein, and Ras-related proteins, which play crucial roles in signaling, apoptosis, stress response, and antigen processing as well as in secretion of danger-associated proteins. These findings collectively indicated that the chicken DCs are expressing various receptors regarded as potential targets for pathogen interaction during viral infection. Therefore, fundamental study of the interaction of DCs and IBDV will provide valuable information in understanding the role of professional antigen-presenting cells in chickens and their molecular interactions during IBDV infection and vaccination.


[The amplification of nucleotide sequences by PCR and the new technics for molecular diagnosis].
The polymerase chain reaction (PCR) is a method for selective amplification of DNA or RNA segments of up to 2 kilobase pairs (kb) or more in length. Repeated cycles of enzymatic primer extension in opposite directions are performed using synthetic oligonucleotides flanking the sequences of interest and the heat-stable DNA polymerase from the archaebacterium Thermus aquaticus (Taq polymerase). The reaction is simple, fast and extremely sensitive, and the DNA or RNA content of a single cell is sufficient to detect a specific sequence. This method has proved useful in simplifying complex analytical protocols in basic and applied molecular biology.


Arias-Stella reaction in nonpregnant women: a clinicopathologic study of nine cases.
The Arias-Stella reaction (ASR) is a common and universally recognized feature of gestational endometrium. That identical cytologic changes may, on occasion, occur outside the endometrium or in nonpregnant women is not as well documented. This report describes the clinical and pathologic findings in nine nonpregnant women whose endometria demonstrated ASR. The defining characteristics of ASR, associated pathologic changes, and differential diagnosis are discussed. All of these women were peri- or postmenopausal, and eight of nine had received exogenous hormones, most (n = 7), progestational agents. ASR did not persist in the four patients who underwent subsequent endometrial examination. None of these patients developed adenocarcinoma, although follow-up was short. ASR is an unusual response to hormonal therapy in nonpregnant patients. It is important to recognize the phenomenon in this setting and not overinterpret the characteristic cytologic atypia as adenocarcinoma.


Reinforcement value and substitutability of sucrose and wheel running: implications for activity anorexia.
Choice between sucrose and wheel-running reinforcement was assessed in two experiments. In the first experiment, ten male Wistar rats were exposed to concurrent VI 30 s VI 30 s schedules of wheel-running and sucrose reinforcement. Sucrose concentration varied across concentrations of 2.5, 7.5, and 12.5%. As concentration increased, more behavior was allocated to sucrose and more reinforcements were obtained from that alternative. Allocation of behavior to wheel running decreased, but obtained wheel-running reinforcement did not change. Overall, the results suggested that food-deprived rats were sensitive to qualitative changes in food supply (sucrose concentration) while continuing to defend a level of physical activity (wheel running). In the second study, 15 female Long Evans rats were exposed to concurrent variable ratio schedules of sucrose and wheel-running, wheel-running and wheel-running, and sucrose and sucrose reinforcement. For each pair of reinforcers, substitutability was assessed by the effect of income-compensated price changes on consumption of the two reinforcers. Results showed that, as expected, sucrose substituted for sucrose and wheel running substituted for wheel running. Wheel running, however, did not substitute for sucrose; but sucrose partially substituted for wheel running. We address the implications of the interrelationships of sucrose and wheel running for an understanding of activity anorexia.


[Results of the surgical treatment of arthro-fibrosis of the knee].
This retrospective study reports the evaluation of the surgical treatment of arthrofibrosis of the knee joint. Patients (n = 53) developed arthrofibrosis due to different index operations and various causes. Based on severe preoperative disability, patients of our series can be considered as difficult to treat. Our management concept suggests a step by step, pathology-oriented treatment: if necessary, arthroscopic arthrolysis and gentle manipulation is supported by posterior capsulotomy. To evaluate the results, we assessed the range of motion of the knee joint, the relative gain in range of motion, extension- and flexion-deficit, as well as the Tegner, Lysholm and Flandry score. As a result of arthrolytic surgery, all 53 patients showed statistically significant (P < 0.01) improvement of the above parameters. The mean gain in the range of motion was 73%. To emphasize the importance of the correct timing of arthrolysis, patients were divided into early and late arthrolysis groups. patients with an early arthrolysis obtained a significantly (P < 0.01) greater improvement in the absolute range of motion than those with a later treatment. By the additional procedure of a posterior capsulotomy, the average extension deficit (absolute) was significantly (P < 0.01) more reduced than for patients without this treatment. Our data support an early therapeutic intervention for patients with a large restriction in the range of motion. Patients with an extension deficit after arthroscopic arthrolysis and gentle manipulation will improve with the additional procedure of posterior capsulotomy.


Fabrication of Decellularized Cartilage-derived Matrix Scaffolds.
Osteochondral defects lack sufficient intrinsic repair capacity to regenerate functionally sound bone and cartilage tissue. To this extent, cartilage research has focused on the development of regenerative scaffolds. This article describes the development of scaffolds that are completely derived from natural cartilage extracellular matrix, coming from an equine donor. Potential applications of the scaffolds include producing allografts for cartilage repair, serving as a scaffold for osteochondral tissue engineering, and providing in vitro models to study tissue formation. By decellularizing the tissue, the donor cells are removed, but many of the natural bioactive cues are thought to be retained. The main advantage of using such a natural scaffold in comparison to a synthetically produced scaffold is that no further functionalization of polymers is required to drive osteochondral tissue regeneration. The cartilage-derived matrix scaffolds can be used for bone and cartilage tissue regeneration in both in vivo and in vitro settings.


Phase I/II trial of preoperative oxaliplatin, docetaxel, and capecitabine with concurrent radiation therapy in localized carcinoma of the esophagus or gastroesophageal junction.
Preoperative chemoradiotherapy is a primary treatment option for patients with resectable esophageal cancer. Combination regimens using newer agents may improve patient outcomes. This multicenter community-based phase I/II trial examined a modern triplet regimen comprised of oxaliplatin, docetaxel, and capecitabine (ODC) combined with radiation therapy (RT). The primary end point was the pathologic complete response (pCR) rate. Eligibility criteria included resectable stage I to III cancer of the mid-/distal-esophagus or gastroesophageal junction, measurable disease, and Eastern Cooperative Oncology Group performance status (ECOG PS) of 0 or 1. Treatment included oxaliplatin 40 mg/m(2), docetaxel 20 mg/m(2) (intravenous, weekly x 5); capecitabine 1,000 mg/m(2) orally twice daily on days 1 to 7, 15 to 21, and 29 to 35; and concurrent RT (45 Gy). Resection was performed during weeks 9 to 12. ODC and RT safety was determined in a phase I portion (n = 10) preceding phase II. Fifty-nine patients were enrolled (September 2005 to February 2008; phase I/cohort 1, 10 patients; phase I/cohort 2/phase II, 49 patients). Baseline characteristics included median age of 63 years; 84% male; ECOG PS 0 and 1, 51% and 49%, respectively; adenocarcinoma and squamous cell, 69% and 18%, respectively; stage I, II, and III, 12%, 41%, and 45%, respectively. Phase I revealed no dose-limiting toxicity. Responses: pCR rate, 49%; objective response rate, 61% (24 complete and six partial responses); stable disease, 6%; and progressive disease, 2%. Sixty-nine percent of patients underwent surgery. Survival: median follow-up, 116 weeks; median disease-free survival (DFS) and overall survival (OS) were 16.3 and 24.1 months, respectively. Two-year DFS and OS were 45.1% and 52.2%, respectively. Most common (>or= 5%) grade 3 to 4 nonhematologic toxicities were anorexia (20%), dehydration (16%), diarrhea (8%), dysphagia (10%), esophagitis (20%), fatigue (12%), hyperglycemia (6%), nausea (16%), pulmonary symptoms (14%), sepsis (6%), and vomiting (16%). All other grade 3 to 4 hematologic and nonhematologic toxicities were uncommon (< 5%). Preoperative ODC plus RT is active and relatively safe in patients with locoregional esophageal cancer. Importantly, this therapy can be administered within 8 weeks. This regimen warrants additional study in this setting and in combination with newer biologic agents.


Use of adult mesenchymal stem cells isolated from bone marrow and blood for somatic cell nuclear transfer in pigs.
Mesenchymal stem cells (MSCs) isolated from bone marrow were used to examine the hypothesis that a less differentiated cell type could increase adult somatic cell nuclear transfer (SCNT) efficiencies in the pig. SCNT embryos were produced using a fusion before activation protocol described previously and the rate at which these developed to the blastocyst stage compared with that using fibroblasts obtained from ear tissue from the same animal. The use of bone marrow MSCs did not increase cleavage rates compared with adult fibroblasts. However, the percentage of embryos that developed to the blastocyst stage was almost doubled, providing support for the hypothesis that a less differentiated cell can increase cloning efficiencies. As MSCs are relatively difficult to isolate from the bone marrow of live animals, a second experiment was undertaken to determine whether MSCs could be isolated from the peripheral circulation and used for SCNT. Blood MSCs were successfully isolated from four of the five pigs sampled. These cells had a similar differentiation capacity and marker profile to those isolated from bone marrow but did not result in increased rates of development. This is the first study to our knowledge, to report that MSCs can be derived from peripheral blood and used for SCNT for any species. These cells can be readily obtained under relatively sterile conditions compared with adult fibroblasts and as such, may provide an alternative cell type for cloning live animals.


Proximal tubule endocytic apparatus as the specific renal uptake mechanism for vitamin D-binding protein/25-(OH)D3 complex.
The renal proximal tubule exhibits a very extensive apical endocytic apparatus that is involved in the reabsorption of molecules filtered in the glomeruli. Several key receptors appear to be involved in this function, which serves not only to conserve protein but also to reabsorb different vitamins in complex with their binding proteins. Recent research has established megalin as probably the most important receptor in this endocytosis process. Cubilin is another receptor identified in the proximal tubule endocytic apparatus. Because cubilin lacks transmembrane or cytoplasmic domains required for endocytosis, this receptor associates with megalin to recycle and internalize its ligands. Recent studies have shown that vitamin D-binding protein (DBP)/25-(OH)D3 complex is one of the megalin/cubilin ligands. Megalin knockout mice develop vitamin D deficiency and bone disease owing to an inability of the proximal tubules to capture the DBP/25-(OH)D3 complexes from the glomerular filtrate. In the same way, kidney-specific megalin knockout mice have severe plasma vitamin D deficiency, hypocalcaemia and serious bone disease, like the complete megalin knockout mice. Anti-cubilin antibodies inhibit cellular uptake of DBP/25-(OH)D3 by up to 70%. Anti-megalin antibodies produced a similar reduction in DBP/25-(OH)D3 endocytosis. When both antibodies were applied, impairment of DBP/25-(OH)D3 was only slightly more impaired (around 80%), suggesting that cubilin and megalin function through the same endocytic pathway. Specific forms of renal Fanconi syndrome are associated with endocytic pathway dysfunction with disruption of megalin-mediated uptake DBP/25-(OH)D3 complex, producing metabolic bone disease in affected individuals as a prominent clinical finding.


Magnetic structure and magnetic properties of synthetic lindgrenite, Cu3(OH)2(MoO4)2.
Synthetic Cu3(OH)2(MoO4)2 consists of Cu3(OH)2 brucite ribbons of edge-sharing copper octahedra connected by MoO4 into a 3D network as in the mineral, lindgrenite, for all temperatures between 1.5 and 300 K. Each ribbon consists of a triangular connection between two different types of copper atom (Cu(1) and 2 Cu(2)) via mu3-OH. The MoO4 acts both as one- and three-atom bridges to connect six Cu atoms belonging to three adjacent ribbons. The magnetic properties are consistent with those of ferrimagnetic chains, and the resulting moment of each chain is parallel below the long-range magnetic ordering at 13 K. The Curie constant is 0.468(1) emu K mol-1 of Cu; the Weiss temperature is -14.2(2) K, and the saturation magnetization at 2 K in 50 kOe is 0.41 N muB mol-1 of Cu. Analyses of the neutron powder diffraction reveal an ordered magnetic state where the moment of Cu(1) is antiparallel to those of the two Cu(2); all of them point along the a axis without any sign of geometrical frustration. Any degeneracy that may be present because of the triangular topology of the Cu atoms (s = 1/2) appears to be lifted by the distortion from an ideal equilateral geometry of the triangle. The entropy, estimated from the heat capacity measurements, attains 50% of the total of 17.7 J K-1 mol-1, close to that expected for three Cu atoms (3R ln 2), up to the long range ordering temperature, and the remaining is associated with the low dimensionality of the material.


Study of the behaviour of lesional and nonlesional skin of canine recurrent flank alopecia transplanted to athymic nude mice.
Recurrent flank alopecia is a clinically well-described skin disorder in dogs. The pathomechanism of the disease is difficult to study because it undergoes spontaneous regression. To study the behaviour of xenografts in mice in order to assess the feasibility of a reproducible experimental model and to investigate local or systemic causes of canine recurrent flank alopecia (cRFA). Skin biopsies were taken from lesional and nonlesional skin of two client-owned dogs with cRFA and grafted onto five athymic mice from a research facility. The lesional skin and xenografted skin were evaluated histologically on day 0 and day 30, respectively. Transplanted lesional and nonlesional canine skin regrew hair within 30 days, while the donor dogs were still alopecic in the lesional areas on day 30 after the skin biopsy procedure. Graft rejection was evidenced histologically in two xenografted athymic mice. Lesional hyperpigmentation disappeared in the athymic mice. This study showed that hair follicles from dogs with cRFA quickly regenerated and regrew hair once grafted onto the mice. Our data indicate that the pathogenesis of cRFA is likely to be mediated by systemic rather than local factors. While this xenograft approach might not be of much value for the study of cRFA, it has potential value for the study of other causes of canine alopecia due to systemic factors.


Feasibility of a curvature-based enhanced display system for detecting cerebral aneurysms in MR angiography.
To assess the feasibility of a curvature-based enhanced display system for detecting cerebral aneurysms in MR angiography. MR angiography studies of 18 patients (eight male and 10 female, average age 65.7, age range 50 to 75 years old) with 23 known aneurysms were evaluated with a curvature-based display system. The two curvature features-the volumetric shape index and curvedness values-were calculated at each voxel. These were displayed independently on a workstation, overlaid on volume-rendered images. Two neuroradiologists evaluated the images for visibility and diagnosis of the cerebral aneurysms. The diagnostic results were compared with the original reports. The calculation time for each curvature index was 30 to 40 s for 120 to 140 slices of original MR angiography data. Shape index images emphasized smooth and round aneurysms more than aneurysms with irregular surfaces. Curvedness images revealed aneurysms well when the aneurysms had diameters that differed from those of the surrounding vessels. The computer-assisted-detection method detected 24 aneurysms, three of which were not pointed out in the initial report. Our results show that the curvature-based display system we have developed is feasible and that it may help to detect small aneurysms that are prone to be overlooked in routine readings.


Ionic diffusion and proton transfer in aqueous solutions of alkali metal salts.
We report on a series of ab initio molecular dynamics investigations on LiCl, NaCl, and KCl aqueous solutions under the effect of static electric fields. We have found that although in low-to-moderate field intensity regimes the well-known sequence of cationic mobilities μ(K+) > μ(Na+) > μ(Li+) (i.e., the bigger the cation the higher the mobility) is recovered, from intense field strengths this intuitive rule is no longer verified. In fact, field-induced water molecular dissociations lead to more complex phenomena regulating the standard migration properties of the simplest monovalent cations. The water dissociation threshold is lowered from 0.35 V Å-1 to 0.25 V Å-1 by the presence of charged species in all samples. However, notwithstanding a one-stage process of water ionization and proton conduction takes place at 0.25 V Å-1 in the electrolyte solutions where "structure maker" cations are present (i.e., LiCl and NaCl), the KCl aqueous solution shows some hindrance in establishing a proton conductive regime, which is characterized by the same proton conduction threshold of neat water (i.e., 0.35 V Å-1). In addition, it turns out that protons flow easily in the LiCl (σp = 3.0 S cm-1) solution and then - in descending order - in the NaCl (σp = 2.5 S cm-1) and KCl (σp = 2.3 S cm-1) electrolyte solutions. The protonic conduction efficiency is thus inversely proportional to the ionic radii of the cations present in the samples. Moreover, Cl- anions act as a sort of "protonic well" for high field intensities, further lowering the overall proton transfer efficiency of the aqueous solutions. As a consequence, all the recorded protonic conductivities are lower than that for neat water (σp = 7.8 S cm-1), which strongly indicates that devices exploiting the proton transfer ability should be designed so as to minimize the presence of ionic impurities.


Air-borne genotype by genotype indirect genetic effects are substantial in the filamentous fungus Aspergillus nidulans.
Genotype by genotype indirect genetic effects (G × G IGEs) occur when the phenotype of an individual is influenced by an interaction between its own genotype and those of neighbour individuals. Little is known regarding the relative importance of G × G IGEs compared with other forms of direct and indirect genetic effects. We quantified the relative importance of IGEs in the filamentous fungus Aspergillus nidulans, a species in which IGEs are likely to be important as air-borne social interactions are known to affect growth. We used a collection of distantly related wild isolates, lab strains and a set of closely related mutation accumulation lines to estimate the contribution of direct and indirect genetic effects on mycelium growth rate, a key fitness component. We found that indirect genetic effects were dominated by G × G IGEs that occurred primarily between a focal genotype and its immediate neighbour within a vertical stack, and these accounted for 11% of phenotypic variation. These results indicate that G × G IGEs may be substantial, at least in some systems, and that the evolutionary importance of these interactions may be underappreciated, especially in microbes. We advocate for a wider use of the IGE framework in both applied (for example, choice of varietal mixtures in plant breeding) and evolutionary genetics (kin selection/kin competition studies).


Catecholamine turnover in rats with ventromedial hypothalamic lesions.
The disappearance of norepinephrine (NE) from brown adipose tissue, heart, and pancreas in vivo has been examined in rats with ventromedial hypothalamic (VMH) lesions and in appropriate control rats. Two weeks after the introduction of these lesions, the rate of NE turnover following blockade of NE synthesis with alpha-methyl-p-tyrosine was significantly greater in brown adipose tissue and heart of rats with VMH lesions than in sham-operated controls. This significant increase occurred whether the animals were fed ad libitum or pair-gained to the sham-operated controls. In the pancreas, on the other hand, no effects of hypothalamic lesions could be detected. When rats were fasted, NE turnover slowed in both sham-operated and VMH-lesioned rats, but the turnover slowed more in sham-operated than in VMH-lesioned rats. When exposed to the cold for 6 h during measurements of NE turnover, VMH-lesioned rats increased their rate of NE turnover as did sham-operated rats. Fasted animals exposed to the cold had significantly higher rates of NE turnover whether or not they also had VMH lesions. Animals with electrolytic or knife-cut VMH lesions showed comparable rates of NE turnover in the fasted state. Eight weeks after hypothalamic lesions, similar studies were conducted. By this time NE turnover was not significantly different between the VMH-lesioned and sham-operated rats fed ad libitum. However, fasting, which slowed NE turnover in sham-operated rats, had no effect in VMH-lesioned animals. These data are discussed in relation to the autonomic hypothesis.


Estrogen does not restore bone lost after ovariectomy in the rat.
We recently found that 17 beta-estradiol (E2) not only suppresses bone resorption but also stimulates bone formation in the cancellous bone of female rats. This raises the possibility that E2 treatment might restore the bone lost after ovariectomy in the rat. To test this, 13-week-old rats were ovariectomized (ox). After a further 13 weeks the animals were injected with E2 (4 mg or 40 micrograms/kg daily), human calcitonin (hCT) (3 IU/kg daily), (3-amino-1-hydroxypropylidene)-1-bisphosphonate (AHPrBP) (0.3 mg/kg twice per week), or a combination of E2 with hCT or AHPrBP, for 8 weeks. The bone volume at the tibial metaphysis of ox animals was approximately 40% of that of sham-operated controls at the end of the experiment. Although the bone volume of ox rats treated with E2 and/or hCT or AHPrBP was slightly higher than that of untreated ox rats, the increase was not significant. Neither E2 alone nor a combination of E2 with hCT or AHPrBP was associated with a higher bone volume than hCT or AHPrBP alone, suggesting no effect of E2 beyond that of inhibition of bone resorption. Histodynamic indices of bone formation were increased in untreated ox rats compared to controls but suppressed in E2-treated, hCT-treated, and AHPrBP-treated animals. These results emphasize the similar responses of rat and human bone, both of which not only show bone loss with estrogen deficiency, preventable by estrogen administration, but also show an inability of estrogen to restore bone lost as a result of estrogen deficiency.


Effect of energy drink and caffeinated beverage consumption on sleep, mood, and performance in children and adolescents.
The increasing availability of highly caffeinated beverages, including energy drinks, in the United States has resulted in a rise in consumption by children and adolescents. In addition, there is mounting evidence that these products are often consumed by youth for their perceived fatigue-mitigating and mood- or performance-enhancing effects. Although such perceptions by children and adolescents about the potential consequences of caffeine consumption are highly likely to influence decision making regarding the use of such products, there is still a relative paucity of studies that focus on the effect of caffeinated beverages on sleep, mood, and performance in the pediatric population. This review summarizes the following aspects of this topic, as derived from the information currently available: 1) the perception, among youth, of caffeine's risks and benefits and the sources of information about caffeine, particularly with regard to sleep, mood, and performance; 2) the bidirectional effect of caffeine on sleep in children and adolescents and the association of caffeine with other sleep-related practices, and 3) the evidence that supports caffeine as a performance and mood enhancer as well as a countermeasure to sleepiness in the pediatric population. Finally, gaps in knowledge are identified, and a direction for future research is outlined.


The Drosophila tumor suppressor gene dlg is required for normal synaptic bouton structure.
The Drosophila tumor suppressor gene lethal (1) discs large (dlg) encodes a protein necessary for normal cell growth in epithelial and brain tissue. It shares high sequence identity to the mammalian synaptic proteins PSD-95 and SAP-70, whose functions are unknown. To determine the localization and role of dlg at synapses, we investigated its distribution and the effects of dlg mutations on Drosophila neuromuscular junctions. We show that dlg immunoreactivity is expressed at one type of glutamatergic synapse and is associated with both presynaptic and postsynaptic membranes. Mutations in dlg alter the expression of dlg and cause striking changes in the structure of the subsynaptic reticulum, a postsynaptic specialization at these synapses. These results indicate that dlg is required for normal synaptic structure and offer insights regarding the role of dlg homologs at vertebrate synapses.


Effect of TEMPO-oxidization and rapid cooling on thermo-structural properties of nanocellulose.
Recently, surface functionality and thermal property of the green nanomaterials have received wide attention in numerous applications. In this study, microcrystalline cellulose (MCC) was used to prepare the nanocrystalline celluloses (NCCs) using acid hydrolysis method. The NCCs was treated with TEMPO [(2,2,6,6-tetramethylpiperidin-1-yl)oxy radical]-oxidation to prepare TEMPO-oxidized NCCs. Cellulose nanofibrils (CNFs) also prepared from MCC using TEMPO-oxidation. The effects of rapid cooling and chemical treatments on the thermo-structural property studies of the prepared nanocelluloses were investigated through FTIR, thermogravimetric analysis-derivative thermogravimetric (TGA-DTG), and XRD. A posteriori knowledge of the FTIR and TGA-DTG analysis revealed that the rapid cooling treatment enhanced the hydrogen bond energy and thermal stability of the TEMPO-oxidized NCC compared to other nanocelluloses. XRD analysis exhibits the effect of rapid cooling on pseudo 2I helical conformation. This was the first investigation performed on the effect of rapid cooling on structural properties of the nanocellulose.


Qualitative and quantitative ultrasound assessment of gastric content.
Pulmonary aspiration of the gastric contents is one of the most feared complications in anesthesia. Its prevention depends on preoperative fasting as well as identification of risky patients. A reliable diagnostic tool to assess gastric volume is currently lacking. The aim of this study performed on volunteers was to evaluate the feasibility of ultrasonography to identify qualitative and quantitative gastric content. A standardized gastric scanning protocol was applied on 67 healthy volunteers to assess the gastric antrum in four different situations: fasting, after ingesting clear fluid, milk and a solid meal. A qualitative and quantitative assessment of the gastric content in the antrum was performed by a blinded sonographer. The antrum was considered either as empty, or containing clear or thick fluid, or solids. Total gastric volume was predicted based on a cross-sectional area of the antrum. A p-value less than 0.05 was considered statistically significant. For each type of gastric content, the sonographic characteristics of the antrum and its content were described and illustrated. Sonographic qualitative assessment allowed to distinguish between an empty stomach and one with different kinds of meal. The predicted gastric volume was significantly larger after the consumption of any food source compared to fasting. Bedside sonography can determine the nature of gastric content. It is also possible to estimate the difference between an empty gastric antrum and one that has some food in it. Such information may be useful to estimate the risk of aspiration, particularly in situations when prandial status is unknown or uncertain.


Extensive Inter-Cyst DNA Methylation Variation in Autosomal Dominant Polycystic Kidney Disease Revealed by Genome Scale Sequencing.
Autosomal dominant polycystic kidney disease (ADPKD) is a heritable disease characterized by bilateral renal enlargement due to the growth of cysts throughout the kidneys. Inheritance of a disease-causing mutation is required to develop ADPKD, which results in end-stage kidney disease and is associated with a high morbidity. The pathology underlying cyst formation is not well understood. To address this, we have previously shown the global methylome is altered in ADPKD tissue, suggesting a role of DNA methylation in disease-state renal tissue. As cysts are believed to arise independently, we hypothesize that DNA methylation changes vary accordingly. Here we further investigate the role of DNA methylation within independent cysts to characterize key intra-individual changes. We demonstrate that fragments within CpG islands and gene bodies harbor the greatest amount of variation across the ADPKD kidney, while intergenic fragments are comparatively stable. A proportion of variably methylated genes were also differentially methylated in ADPKD tissue. Our data provide evidence that individual molecular mechanisms are operating in the development of each cyst.


Effect of dietary supplementation with yeast cell wall extracts on performance and gut response in broiler chickens.
The dietary supplementation of yeast cell wall extracts (YCW) has been found to reduce pathogenic bacteria load, promote immunoglobulin production, prevent diseases by pro-inflammatory responses, and alter gut microbiota composition. This study evaluated growth and slaughter results, health, gut morphology, immune status and gut transcriptome of 576 male chickens fed two diets, i.e. C (control) or Y (with 250-500 g/t of YCW fractions according to the growth period). At 21 and 42 d the jejunum of 12 chickens per diet were sampled and stained with hematoxylin/eosin for morphometric evaluation, with Alcian-PAS for goblet cells, and antibodies against CD3+ intraepithelial T-cells and CD45+ intraepithelial leukocytes. The jejunum sampled at 42 d were also used for whole-transcriptome profiling. Dietary YCW supplementation did not affect final live weight, whereas it decreased feed intake (114 to 111 g/d; P ≤ 0.10) and improved feed conversion (1.74 to 1.70; P ≤ 0.01). Regarding the gut, YCW supplementation tended to increase villi height (P = 0.07); it also increased the number of goblet cells and reduced the density of CD45+ cells compared to diet C (P < 0.001). In the gut transcriptome, four genes were expressed more in broilers fed diet Y compared to diet C, i.e. cytochrome P450, family 2, subfamily C, polypeptide 23b (CYP2C23B), tetratricopeptide repeat domain 9 (TTC9), basic helix-loop-helix family member e41 (BHLHE41), and the metalloreductase STEAP4. Only one gene set (HES_PATHWAY) was significantly enriched among the transcripts more expressed in broilers fed diet Y. However, a total of 41 gene sets were significantly over-represented among genes up-regulated in control broilers. Notably, several enriched gene sets are implicated in immune functions and related to NF-κB signaling, apoptosis, and interferon signals. The dietary YCW supplementation improved broiler growth performance, increased gut glycoconjugate secretion and reduced the inflammatory status together with differences in the gut transcriptome, which can be considered useful to improve animal welfare and health under the challenging conditions of intensive rearing systems in broiler chickens.


Muscle fibre recruitment and metabolism in prolonged exhaustive dynamic exercise.
The rather constant amount of glycogen found in all fibre types in human skeletal muscle provides an opportunity to study the pattern of glycogen depletion with exercise, which should give an indication of which fibres are activated to generate the force. In very light dynamic contractions repeated for hours there is a primary reliance on slow twitch (ST) fibres with no or very minor involvement of fast twitch (FT) fibres. At heavier work loads (greater than 50% Vo2max) ST fibres are depleted first but FT fibres begin to become depleted. Exhaustion at these work levels coincides with muscle fibres of all types being depleted of glycogen. The crucial role of muscle glycogen in both the metabolic response to exercise and work performance is apparent. It is more difficult to explain why extramuscular substrates (plasma free fatty acids) cannot be utilized at a high enough rate to accommodate the energy turnover needed in more intense dynamic exercise. A limitation on the uptake of free fatty acids by the muscle cell rather than its transport to the cell or oxidation within it appears to be the critical factor.


Increased membrane lipid peroxidation and decreased Na+/K+-ATPase activity in erythrocytes of patients with stable coronary artery disease.
We aimed to determine erythrocyte membrane lipid peroxidation levels and Na/K-ATPase activity in patients with and without coronary artery disease (CAD) documented by coronary angiography. A total of 144 patients who had undergone coronary angiography were divided into a CAD group (n=82) and a non-CAD group (control group, n=62) according to the results of coronary angiography. Lipid peroxide levels in plasma and the erythrocyte membrane were measured using a fluorimetric method. Total antioxidant status and Na/K-ATPase activity in plasma were determined using spectrophotometric methods. Lipid peroxidation levels were significantly higher in the erythrocyte membrane of CAD patients compared with controls, whereas Na/K-ATPase activity was significantly lower in the erythrocyte membrane of CAD patients compared with controls. The coronary artery scores were correlated positively with membrane lipid peroxidation (r=0.324, P<0.001) and negatively with Na/K-ATPase activity (r=-0.302, P<0.001). This study shows that the levels of membrane lipid peroxidation and Na/K-ATPase activity are correlated with the severity of CAD.


Uterine giant cell tumor of soft tissue: A case report.
Giant cell tumor of soft tissue (GCT-ST) is a rare primary soft tissue tumor. GCT-ST mainly occurs in the trunk and extremities. There is no standard treatment for GCT-ST. This paper reports a rare case of primary uterine GCT-ST. A 48-year-old female patient underwent a transabdominal subhysterectomy for uterine leiomyoma. Postoperative pathological examination showed GCT-ST with unclear tissue boundary (10.0 × 6.0 × 5.0 cm). A small amount of GCT-ST tissue could be seen on the local edge of the leiomyoma. Residual tumor tissue was found around the uterine cavity. The patient reported persistent lower abdominal distension pain 3 months after the operation. Pelvic and abdominal imaging showed a huge tumor and multiple pelvic and abdominal organ metastasis. No pulmonary metastasis was found. Exploratory surgery revealed widespread metastases in the abdominal and peritoneal cavities, involving both ovaries, right tubal serous membrane, appendix serous membrane, bladder, pelvic peritoneum, and abdominal wall incision. After surgery, the patient had 6 cycles of docetaxel and carboplatin but stopped treatments due to economic reasons. The patient died 3 months later because of multiple organs failure. GCT-ST is generally benign but has unpredictable behavior. A massive recurrence with wide invasion is possible after subtotal resection.


Prenatal diagnosis of placenta accreta by colour Doppler ultrasonography: 5-year review.
To determine the accuracy of colour Doppler ultrasonography to diagnose placenta accreta. The authors reviewed cases of placenta accreta between January, 2008 and December, 2012. Ultrasonographic images consistent with signs ofplacenta accreta (numerous vascular lacunae, loss ofsubplacentalsonolucent space, absent lower uterine segment between bladder-placenta, turbulent or complicated blood flow at the uteroplacental interface) were correlated with findings at the time of surgery and pathologic examination. Over 60 months, 12 cases (0.48/1,000 deliveries) with suspected placenta accreta by ultrasonography were studied. The median gestational age atfirst diagnosis was 24 weeks. All cases had at least one previous cesarean delivery. At surgery, all cases had an adherent placenta requiring hysterectomy (five accreta, three increta, andfourpercreta). Four cases (33%) had accidental tear of urinary bladder Nine cases (75%) required blood transfusions. Colour Doppler ultrasonography appears useful in antenatal diagnosis ofplacenta accreta.


Pregnancy-related changes in the longer-term management of HIV-infected women in Europe.
To clarify policies regarding management of HIV-infected women, relating to pregnancy, in view of current European consensus guidelines. Postal questionnaire survey in 36 hospitals in 11 European countries. Responses were received from 22 (61%) centres, representing all 11 countries. In principle, antiretroviral therapy (ART) would be reviewed in treated women wanting to become pregnant in nearly all centres. Multidisciplinary management of infected pregnant women was routine in 17 (77%) centres, facilitating continuity of care. Approximately half the clinicians would use zidovudine monotherapy for pregnant women without indications for ART, while the remainder prescribed combination therapy. In 1998-2000, pre-eclampsia was the most prominent adverse event (22 centres) in women receiving ART, with congenital abnormalities (13 abnormalities in 6 centres) and stillbirth (5 centres) also reported. Policies varied, particularly regarding prophylactic ART for women without indications for treatment and did not always follow current European guidelines.


Cutting-Edge Technologies for Inflamed Joints on Chip: How Close Are We?
Osteoarthritis (OA) is a painful and disabling musculoskeletal disorder, with a large impact on the global population, resulting in several limitations on daily activities. In OA, inflammation is frequent and mainly controlled through inflammatory cytokines released by immune cells. These outbalanced inflammatory cytokines cause cartilage extracellular matrix (ECM) degradation and possible growth of neuronal fibers into subchondral bone triggering pain. Even though pain is the major symptom of musculoskeletal diseases, there are still no effective treatments to counteract it and the mechanisms behind these pathologies are not fully understood. Thus, there is an urgent need to establish reliable models for assessing the molecular mechanisms and consequently new therapeutic targets. Models have been established to support this research field by providing reliable tools to replicate the joint tissue in vitro. Studies firstly started with simple 2D culture setups, followed by 3D culture focusing mainly on cell-cell interactions to mimic healthy and inflamed cartilage. Cellular approaches were improved by scaffold-based strategies to enhance cell-matrix interactions as well as contribute to developing mechanically more stable in vitro models. The progression of the cartilage tissue engineering would then profit from the integration of 3D bioprinting technologies as these provide 3D constructs with versatile structural arrangements of the 3D constructs. The upgrade of the available tools with dynamic conditions was then achieved using bioreactors and fluid systems. Finally, the organ-on-a-chip encloses all the state of the art on cartilage tissue engineering by incorporation of different microenvironments, cells and stimuli and pave the way to potentially simulate crucial biological, chemical, and mechanical features of arthritic joint. In this review, we describe the several available tools ranging from simple cartilage pellets to complex organ-on-a-chip platforms, including 3D tissue-engineered constructs and bioprinting tools. Moreover, we provide a fruitful discussion on the possible upgrades to enhance the in vitro systems making them more robust regarding the physiological and pathological modeling of the joint tissue/OA.


Development of visual-evoked potentials to radially modulated concentric patterns.
The visual processing of radially modulated concentric patterns was studied in human participants, aged 3-22 years, by recording event-related potentials. These stimuli are known to activate the fusiform face area as well as area V4 in normal adults. The electrophysiological data showed a P1 latency that reached a maturation asymptote before 3 years of age, whereas that of N1 and P2 became adultlike by 13 years of age. In addition, the distribution of the P2 component over the scalp was focalized in the primary visual cortex before adolescence and became distributed over the entire brain after adolescence. Radially modulated concentric stimuli thus induce brain activation that is not mature until 13 years of age.


[Expert consensus on the diagnosis and management of Birt-Hogg-Dubé syndrome].
Birt-Hogg-Dubé syndrome (BHD) is a rare autosomal dominant disorder characterized by diffuse pulmonary cysts often leading to recurrent spontaneous pneumothorax, cutaneous fibrofolliculomas or trichodiscomas, and a variety of renal cell cancers. It is caused by pathogenic variants in the FLCN gene located on chromosome 17p11.2. Although an increasing number of patients with BHD syndrome are being recognized in China, the missed diagnosis and delayed diagnosis are still common. In addition, appropriate management is difficult for most of them. Pulmonary cysts and pneumothorax are the main presenting features, but skin and renal lesions appear to be less common in Chinese subjects than those reported from European and American countries. Therefore, the consensus is established by experts from the related disciplines to improve the diagnosis and management of BHD syndrome. This consensus consists of 15 recommendations related to BHD syndrome, including clinical assessments, diagnosis, differential diagnosis, treatment, follow-up, and family management. In particular, it provides revised diagnostic criteria based on the Chinese situation. We hope to promote scientific and clinical progress in this rare disease and improve the prognosis of the patients.Summary of recommendations[Recommendation 1] The folliculin (FLCN) gene is currently the only affirmative causative gene for Birt-Hogg-Dubé (BHD) syndrome, and the pedigree analysis of genetic testing of family members' samples can assist in the rapid identification of causative gene variants. The genetic testing methods, including Sanger sequencing, Multiplex Ligation-dependent Probe Amplification (MLPA), and Next-Generation Sequencing (NGS), can be chosen based on individual patient's care needs. (43 voting experts; 43 in favor, 0 against, 0 abstention).[Recommendation 2] Patients with BHD syndrome should undergo chest CT scan to evaluate cystic lesions in the lungs, and routine evaluation of the kidneys for tumor foci, using ultrasound, enhanced CT, or MR as appropriate. (43 experts voted; 43 in favor, 0 against, 0 abstention).[Recommendation 3] Clinicians should establish a diagnosis based on the appropriate clinical presentation and in conjunction with genetic test results and/or a family history of BHD syndrome. (43 voting experts; 43 in favor, 0 against, 0 abstention).[Recommendation 4] Lung histopathological biopsy is not recommended as the first choice for patients with suspected BHD syndrome based on clinical and pulmonary imaging manifestations. (43 voting experts; 43 in favor, 0 against, 0 abstention).[Recommendation 5] BHD syndrome should be particularly distinguished from other diffuse cystic lung diseases, such as lymphangioleiomyomatosis (LAM), lymphocyte interstitial pneumonia (LIP), pulmonary Langerhans cell histiocytosis (PLCH), etc. (43 voting experts; 43 in favor, 0 against, 0 abstention).[Recommendation 6] BHD syndrome is clinically rare and often involves multiple disciplines, such as respiratory and critical care medicine, radiology, pathology, thoracic surgery, urology, genetics, and dermatology, and multidisciplinary discussions are recommended to improve the diagnosis of BHD syndrome. (43 voting experts; 43 in favor, 0 against, 0 abstentions).[Recommendation 7] Patients with BHD syndrome should avoid smoking, and are recommended to be vaccinated with influenza, pneumococcal, and SARS-Cov-2 vaccines to prevent infections. (43 voting experts; 43 in favor, 0 against, 0 abstentions).[Recommendation 8] Air travel is not recommended for patients with BHD syndrome who have experienced pneumothorax until it has been recovered. (43 voting experts; 42 in favor, 0 against, 1 abstention).[Recommendation 9] In patients with BHD syndrome complicated by pneumothorax, early pleurodesis is recommended to reduce the risk of recurrence. (43 voting experts; 43 in favor, 0 against, 0 abstention).[Recommendation 10] Observation or topical treatment may be chosen for patients with BHD syndrome complicated by fibrofolliculoma or trichodiscoma. (43 voting experts; 42 in favor, 0 against, 1 abstention).[Recommendation 11] For patients with BHD syndrome complicated by renal tumors, annual abdominal MR examination is recommended when tumors are <1 cm in diameter; when tumors are 1-3 cm in diameter, abdominal MR examination at every 6 months or ablation surgery is recommended; when renal tumors are >3 cm in diameter, local excision of renal tumors with preservation of renal function is recommended. (43 voting experts; 43 in favor, 0 against, 0 abstention).[Recommendation 12] Patients with BHD syndrome are associated with significantly increased risks of kidney cancer, and routine screenings of kidney cancer during their lifetime are recommended. (43 voting experts; 43 in favor, 0 against, 0 abstention).[Recommendation 13] Couples with BHD syndrome are advised to undergo prenatal genetic counseling when preparing for pregnancy, to work with a prenatal diagnostician to assess genetic risk, and to discuss the feasibility of prenatal diagnosis during pregnancy. (43 experts voted; 43 in favor, 0 against, 0 abstention).[Recommendation 14] Family members of patients with BHD syndrome should receive health education, and FLCN gene testing is recommended for asymptomatic adults to rule out BHD syndrome in a timely manner. (43 voting experts; 43 in favor, 0 against, 0 abstention).[Recommendation 15] Improvements in the clinical diagnosis and treatment of BHD syndrome and overall management are needed. Due to the scarcity of effective therapeutic drugs, multicenter, prospective clinical trials are recommended. (43 experts voted; 43 in favor, 0 against, 0 abstention).


Professional psychology in Germany, National Socialism, and the Second World War.
Hundreds of positions for psychologists were established after the National Socialists seized power in 1933. It has accordingly been asserted that professional psychology in Germany experienced significant growth during the National Socialist period. An analysis of archival materials and of a recent collection of biographies indicates otherwise, however. German psychology, in fact, declined because of systematic persecution and a surge of emigration, a deficit that subsequent cohorts were barely able to make up until 1945. The new positions for psychologists were mainly in the military testing service, and could only be filled after manpower was shifted from the civilian to the military sector. In 1941, the Ministry of Science and Education released regulations for an innovative practice-oriented national curriculum for psychology. The move was in line with Nazi policy, but it was initiated by a group of protagonists from psychology under the aegis of the German Psychological Association, not the National Socialist German Workers Party, the government, or the military. The present article elaborates how the conception of practice-oriented, state-approved studies was part of the traditional German dual system of academic and professional qualification, and thus actually predated 1933. The new curriculum was largely not implemented because of the exigencies of the war. However, as a regulatory framework it marked a turning point in the merging of academic and professional psychology in Germany. The relationship between academic and professional psychology is also discussed, along with the role of German psychology vis-à-vis National Socialism and the German military. (PsycINFO Database Record


Interactome: gateway into systems biology.
Protein-protein interactions are fundamental to all biological processes, and a comprehensive determination of all protein-protein interactions that can take place in an organism provides a framework for understanding biology as an integrated system. The availability of genome-scale sets of cloned open reading frames has facilitated systematic efforts at creating proteome-scale data sets of protein-protein interactions, which are represented as complex networks or 'interactome' maps. Protein-protein interaction mapping projects that follow stringent criteria, coupled with experimental validation in orthogonal systems, provide high-confidence data sets immanently useful for interrogating developmental and disease mechanisms at a system level as well as elucidating individual protein function and interactome network topology. Although far from complete, currently available maps provide insight into how biochemical properties of proteins and protein complexes are integrated into biological systems. Such maps are also a useful resource to predict the function(s) of thousands of genes.


Plaque removal and oral health promotion potential for the elmex interX medium toothbrush: clinical efficacy and safety evaluation.
This study was designed to assess a newly developed toothbrush for efficacy in plaque removal, reduction of gingivitis and soft tissue bleeding, and retardation of stain formation over 30 days. Healthy adult subjects (N = 112) were admitted to the study, and randomly given a test toothbrush, (the elmex interX medium) or the control toothbrush, an ADA-Accepted flat-bristle profile toothbrush. Additionally, the subjects were randomly assigned to be given oral hygiene instruction, or no formal oral hygiene instruction. Examinations and assessments were performed at baseline, 15 and 30 days, and included gingivitis levels, stain levels, bleeding on probing, and plaque levels before and after brushing. No significant statistical differences were found between the two toothbrushes on gingivitis, nor on the stain index; however, over the course of the study the gingivitis decreased significantly (p = 0.012) for the test toothbrush group. For the bleeding index, a significant statistical difference (p = 0.027) was found at 30 days favoring the test toothbrush. There was also a significant decrease for the test toothbrush on bleeding over time (p = 0.004). Following overnight plaque formation, the before-brushing plaque levels were significantly different in favor of the test toothbrush at 15 days (p = 0.002), and again at 30 days (p = 0.001). Immediately after brushing, there was a significant statistical difference (p = 0.015) favoring the test toothbrush at 30 days. These results clearly indicate improved plaque removal and reduction of bleeding and gingivitis for the new elmex interX medium toothbrush compared to the ADA-Accepted control toothbrush.


Prediction and classification of Alzheimer disease based on quantification of MRI deformation.
Detecting early morphological changes in the brain and making early diagnosis are important for Alzheimer's disease (AD). High resolution magnetic resonance imaging can be used to help diagnosis and prediction of the disease. In this paper, we proposed a machine learning method to discriminate patients with AD or mild cognitive impairment (MCI) from healthy elderly and to predict the AD conversion in MCI patients by computing and analyzing the regional morphological differences of brain between groups. Distance between each pair of subjects was quantified from a symmetric diffeomorphic registration, followed by an embedding algorithm and a learning approach for classification. The proposed method obtained accuracy of 96.5% in differentiating mild AD from healthy elderly with the whole-brain gray matter or temporal lobe as region of interest (ROI), 91.74% in differentiating progressive MCI from healthy elderly and 88.99% in classifying progressive MCI versus stable MCI with amygdala or hippocampus as ROI. This deformation-based method has made full use of the pair-wise macroscopic shape difference between groups and consequently increased the power for discrimination.


[Molecular techniques applied in species identification of Toxocara].
Toxocarosis is still an important and actual problem in human medicine. It can manifest as visceral (VLM), ocular (OLM) or covert (CT) larva migrans syndroms. Complicated life cycle of Toxocara, lack of easy and practical methods of species differentiation of the adult nematode and embarrassing in recognition of the infection in definitive hosts create difficulties in fighting with the infection. Although studies on human toxocarosis have been continued for over 50 years there is no conclusive answer, which of species--T. canis or T. cati constitutes a greater risk of transmission of the nematode to man. Neither blood serological examinations nor microscopic observations of the morphological features of the nematode give the satisfied answer on the question. Since the 90-ths molecular methods were developed for species identification and became useful tools being widely applied in parasitological diagnosis. This paper cover the survey of methods of DNA analyses used for identification of Toxocara species. The review may be helpful for researchers focused on Toxocara and toxocarosis as well as on detection of new species. The following techniques are described: PCR (Polymerase Chain Reaction), RFLP (Restriction Fragment Length Polymorphism), RAPD (Random Amplified Polymorphic DNA) and SSCP (Single Strand Conformation Polymorphism).


[Features of the biosynthesis of immunoglobulin G peculiar to the growth process].
It is established that with partial hepatectomy, Shvets leukosis and hepatoma RS-1 the biosynthesis intensity of rat blood serum proteins producing aggregates in the acid medium is considerably higher than that of other serum proteins, the incorporation of the radioactive precursor into immunoglobulin G peculiar to intensive normal and malignant growth being particularly intensive. During liver regeneration as well as in malignant growth specific radioactivity of immunoglobulin G peculiar to the growth processes is three and five times, respectively, as high as this value for blood serum soluble proteins and proteins of alpha-globulin fractions.


Rectus and oblique muscles of eyeball: a morphometric study of Indian population.
During the strengthening and weakening procedures of intraocular muscles, distance of insertion from the sclerocorneal junction is an important determinant in the identification of muscles. During repositioning of the aponeurosis of the muscles, it is desired that the width should not change in order to avoid diversion of forces. Available anatomic studies on insertions of extraocular muscles are few, date back to early twentieth century and have been conducted on mostly white population. The present study is an attempt to document the insertions of recti and oblique muscles in Indian population. Forty eyeballs were removed from orbit. Insertion of recti and obliqui were cleaned and eyeballs were perfused with normal saline to regain the volume (hence shape and size) before recording observations. Insertion of recti and obliqui muscles were observed under various study parameters. The distance of insertion of recti from the limbus were found to be 7.3 mm, 8.06 mm, 8.71 mm, and 8.74 mm for medial, inferior, lateral, and superior rectus, respectively. The superior oblique was aponeurotic and found to be more variable in mode of insertion as compared to inferior oblique which had a fleshy and relatively constant insertion. The observations on insertion of recti and obliqui as obtained in present study differ from earlier studies to the tune of 1-1.5 mm. This may be attributed to adoption of method of reperfusion of eyeball before recording observations thus maintaining size close to in vivo. The observations are expected to be closer to actual.


Multiple pH levels in chromatograms.
Lines of buffer solutions drawn longitudinally on chromatograms with a capillary pipette provide different levels of pH which affect the Rf values, color, fluorescence, and other properties of the chromatographed substances.


Assessment of apoptosis by immunohistochemistry to active caspase-3, active caspase-7, or cleaved PARP in monolayer cells and spheroid and subcutaneous xenografts of human carcinoma.
Immunohistochemistry to active caspase-3, recently recommended for apoptosis detection, is inappropriate to detect apoptosis involving caspase-7. Cleavage of poly-ADP-ribose polymerase 1 (PARP-1), a major substrate of both caspases, is a valuable marker of apoptosis. Apoptosis evaluation induced in vitro either by paclitaxel or by photodynamic treatment (PDT) with Foscan in HT29 or KB monolayer cells and HT29 spheroids yielded a close percentage of labeled cells whatever the antibody used, whereas in control specimens, cleaved PARP (c-PARP) immunostaining failed to detect apoptosis as efficiently as active caspase-3 or -7 immunostaining. Studies in MDA-MB231 monolayer cells and HT29 xenografts either subjected or not subjected to Foscan-PDT resulted in a significant higher number of active caspase-3-labeled cells, although immunofluorescence analysis showed c-PARP and active caspase-3 perfectly colocalized in tumors. A restricted expression of c-PARP was obvious in the greater part of caspase-3 expressing cells from control tumor, whereas photosensitized tumors showed a higher number of cells expressing large fluorescent spots from both active caspase-3 and c-PARP. These results support the assumption that c-PARP expression was dependent on treatment-induced apoptosis. The absence of caspase-7 activation in some caspase-3-expressing cells undergoing Foscan-PDT shows the relevance of using antibodies that can discriminate caspase-dependent apoptotic pathways.


Hairpins in bookstacks: information retrieval from biomedical text.
Current advances in high-throughput biology are accompanied by a tremendous increase in the number of related publications. Much biomedical information is reported in the vast amount of literature. The ability to rapidly and effectively survey the literature is necessary for both the design and the interpretation of large-scale experiments, and for curation of structured biomedical knowledge in public databases. Given the millions of published documents, the field of information retrieval, which is concerned with the automatic identification of relevant documents from large text collections, has much to offer. This paper introduces the basics of information retrieval, discusses its applications in biomedicine, and presents traditional and non-traditional ways in which it can be used.


Intermediate filament proteins participate in signal transduction.
How timely transport of chemical signals between the distal end of long axonal processes and the cell bodies of neurons occurs is an interesting and unresolved issue. Recently, Perlson et al. presented evidence that cleavage products of newly synthesized vimentin, an intermediate filament (IF) protein, interact with mitogen-activated protein (MAP) kinases at sites of axon injury. These IF fragments appear to be required for the transport of these kinases to the cell body along microtubule tracks. The truncated vimentin is instrumental in signal propagation as it provides a scaffold that brings together activated MAP kinases (such as Erk 1 and Erk2), as well as importin beta and cytoplasmic dynein. The authors propose that this all-in-one transport complex has the extraordinary ability to travel towards the cell body and enter the nucleus where the kinases activate and influence gene expression so that a neuron can generate a timely response to injury.


Identification of a T-cell epitope adjacent to neutralization antigenic site 1 of poliovirus type 1.
Proliferative T-cell responses to poliovirus in various strains of mice have been analyzed by using either killed purified virus or capsid protein VP1 synthetic peptides. Following immunization of mice with inactivated poliovirus type 1 (PV1), a specific proliferative response of their lymph node CD4+ T cells was obtained after in vitro stimulation with purified virus. In mice immunized with PV1, PV2, or PV3, a strong cross-reactivity of the T-cell responses was observed after in vitro stimulation with heterologous viruses. By using various strategies, a dominant T-cell epitope was identified in the amino acid 103 to 115 region of capsid polypeptide VP1, close by the C3 neutralization epitope. The T-cell response to VP1 amino acids 103 to 115 is H-2 restricted: H-2d mice are responders, whereas H-2k and H-2b mice do not respond to this T-cell epitope. Immunization of BALB/c (H-2d) mice with the uncoupled p86-115 peptide, which represents VP1 amino acids 86 to 115 and contains both the T-cell epitope and the C3 neutralization epitope, induced poliovirus-specific B- and T-cell responses. Moreover, these mice developed poliovirus neutralizing antibodies.


Contents of Ag and other metals in food-contact plastics with nanosilver or Ag ion and their migration into food simulants.
Six nanosilver-labelled products and five silver ion (Ag(+))-labelled products were investigated to measure the migration of Ag from food-contact plastics, including nanosilver into various food simulants. The products were obtained in Japanese markets in 2012. Zinc (Zn), another major antimicrobial agent, and three harmful metals, cadmium (Cd), lead (Pb) and arsenic (As), were also examined. Ag and Zn were detected in all six nanosilver products at concentrations of 21-200 and 8.4-140 mg kg(-1), respectively. These metals were also detected in all five Ag(+) products at the same level as nanosilver products. Cd, Pb and As were not detected in any sample. Migrations of Ag and Zn were highest in 4% acetic acid, but also observed in water and 20% ethanol. Big differences were not observed in the migration ratio between nanosilver products and Ag(+) products. The ultrafiltration experiments suggested that the Ag that migrated from nanosilver products into 4% acetic acid was in its ionic form, while that into water and 20% ethanol was in its nanoparticle form.


A Pilot Study of the Short Term Effectiveness and Safety of Amniotic Fluid in Severe Dry Eye Disease.
The aim of this study was to evaluate if amniotic fluid (AF) mixed with artificial tears or soaked with a soft contact lens (SCL) as a treatment for severe dry eye disease (DED) would improve its signs or symptoms. In this retrospective pilot study 22 consecutive eyes of 11 patients with level 3 DED classified by DEWS 1 (Dry Eye WorkShop 1 2007), were included in the study between June 1 and September 30 in 2017. The study was conducted before DEWS II (Dry Eye WorkShop II 2017), which was published in October 2017. Therefore, DEWS II was not adopted for this study. Soft Contact Lens Acuvue Oasys of Plano with 8.8 base curve and 14 mm diameters by Johnson and Johnson were used to soak in FloGraft, which is an AF, for 30 minutes before placing in 12 eyes in Group 1. The contact lenses were placed in the left eye for 1 week. In Group 2, 10 eyes used 6 mL of artificial tears mixed with 0.25 mL of AF, which were applied to the eye four times a day for 1 week. No other eye medications were used. The eyes that were included had diffuse punctate staining and fast tear breaking times of <5 seconds with severe ocular symptoms (DEWS 1 level 3-4). Photos of fluorescein stain corneas before the treatment and 1 week after the treatment were used to compare the distribution of punctate staining as the objective outcomes for signs. Several questions adopted from the Ocular Surface disease Index (OSDI) about subjective symptoms before and after the treatment were asked, and documented on the chart. Improvement either in symptoms or signs or both of DED after 1 week at follow-up examination was recorded. Group 1, with SCL 46% had improvement after 1 week and Group 2, with artificial tears 50% had improvement after 1 week. Improvement means either symptom relief or comparatively decreased distribution of punctate staining on the cornea or both. No cases had inflammation, infection, irritation or blurred vision. We concluded that FloGraft as an AF can safely improve the symptoms or the signs of severe DED either as a mix with artificial tears or soaked with SCL by about 50% in this case series without evidence of irritation, inflammation or blurred vision in the short term.


[Role of surgery in the treatment of pulmonary aspergillosis].
This article reviews the different forms of pulmonary disease caused by aspergillus and discusses the possible surgical treatments. The most well known is the classic aspergilloma which develops as a fungal ball in the centre of a pre-existing pulmonary cavity. One can distinguish simple (few symptoms, thin walled cavity without immediate complications) and complex forms (patient generally unwell, thick cavity, complications). In the complex form, surgical intervention must be considered as a last resort. In the simple form, surgery is relatively benign and prevents disease progression. Pleural aspergillosis can occur, usually following the surgical removal of a cavity either in the short or medium term. Given the loss of lung parenchyma thoracoplasty is often the only option. Two different scenarios occur in acute invasive aspergillosis where surgery may be indicated: firstly, surgery can be considered in the event of haemoptysis related to vascular erosion; secondly, resection of mycotic sequestrations before intensification or resumption of therapy may prevent a relapse. Semi-invasive aspergillosis usually occurs in territories of post-radiation fibrosis: after a phase of invasion equivalent to a lobar pneumonia, a secondary cavity appears containing a small fungal ball. Thoracoplasty is often the only surgical option. Ulcerating tracheobronchial aspergillosis has been observed following (cardio-) pulmonary transplant and this may progress to a characteristic invasive aspergillosis. Finally, rare observations of parietal aspergillosis could be treated by surgical resection and associated with systemic antifungal therapy. Optimum management of these patients requires a multidisciplinary approach.


Differentiation of crypt epithelium in human palatine tonsils: the microenvironment of crypt epithelium as a lymphoepithelial organ.
The differentiation of the keratinocytes of the human palatine tonsils were studied by means of light and electron microscopy and immunohistochemistry using a polyclonal (K) and two monoclonal antikeratin antibodies (PKK1, PKK2). In the surface epithelium, the basal cells, cuboidal or columnar in shape, undergo progressive terminal differentiation and are transformed into the flattened cells of the upper layers. K reacts with both the basal and spinous layers, while PKK1 and PKK2 mark exclusively the basal layer. In the neck portion of the crypt, cavities containing one or aggregated lymphocytes with amorphous substances are observed in the spinous layer. The cavities are surrounded by elongated cytoplasmic processes of transformed epithelial cells bearing surface microvilli. These transformed epithelial cells display intense PKK1- and PKK2-positive reactions, whereas other conventional polygonal cells in the vicinity remain PKK1- and PKK2-negative as do those in the surface epithelium. In the deep portion of the crypt, where numerous lymphocytes invade the epithelium, the epithelial cells are transformed into star-shaped reticulum cells showing PKK1- and PKK2-positive reactions. The extended and branched cytoplasmic processes interconnect with one another constituting a complex network of reticulum cells, the well known reticulation of the crypt epithelium. Ten-nm filaments are usually oriented parallel to the longitudinal axis of transformed epithelial cells. Our observations suggest that the cell-shape transformation, i.e., from conventional polygonal epithelial cells into epithelial reticulum cells, occurs when the epithelial cells are in close contact with the infiltrating lymphocytes, and that this transformation is accompanied by a change in keratin phenotype.


[Common questions and suggestions of evaluation for NDA of TCM].
According to the existing Provisions for Drug Registration (SFDA Order No. 28), applications for new drugs of traditional Chinese medicine are divided into two parts: the applications for drug clinical trial and for drug production (including new drug certificate). It will last for about 10 years from the application for drug clinical trial to get approving, and it also remains many problems and the low probability to succeed. From the sight of pharmaceutical review, there are mainly two aspects of regulatory compliance and technical issues, mainly for changes without approval of the competent authorities of the country. For example, sample preparation and approval of clinical trial process are significant changes. Technical problems are reporting incomplete data or information submitted does not comply with the technical requirements for review, such as: production process validation does not provide information, the preparation of samples for clinical trials and field inspection, production information, or the information provided does not meet the technical requirements. This paper summarizes the frequently asked questions and to make recommendations to advise applicants concerned, timely detection of problems, avoid risk, improving the quality and efficiency of the application for registration.


Measurements of single event spectra with a wall-less proportional counter in low LET radiation fields.
Measurements have been made on 60Co gamma rays from a point source and on 6 MV roentgen rays from a linear accelerator with a wall-less proportional counter. Single event spectra have been measured in sites of 0.5-2 mum in diameters stimulated with a tissue equivalent gas. Calculations of the relative variance in LET are performed and compared with the relative variance of the measured distributions for estimates about the straggling contribution.


The Comparative Effectiveness of Monotherapy and Combination Therapies: Impact of Angiotensin Receptor Blockers on the Onset of Alzheimer's Disease.
The criteria for use of Alzheimer's disease (AD) drug Leqembi recommended by the Department of Veterans Affairs (VA) include patients aged 65 years or older with mild cognitive impairment (MCI) or mild AD. Comorbidities that include hypertension, hyperlipidemia, and diabetes are common among these patients. Our objective is to investigate the comparative effectiveness of the administration of one, two, or three medications belonging to the categories of angiotensin receptor blockers (ARBs), angiotensin-converting enzyme inhibitors (ACEIs), Beta Blockers, Statins, and Metformin, for their potential to delay the clinical onset of AD and provide a window of opportunity for therapeutic intervention. Retrospective matched case-control study. Data from the Department of Veterans Affairs national corporate data warehouse. We conducted an analysis of 122,351 participants (13,611 with AD and 108,740 without AD), aged 65-89, who began at least one of the prescribed medication classes under investigation between October 1998 and April 2018. We utilized Cox proportional hazard regressions, both with and without propensity score weighting, to estimate hazard ratios (HR) associated with the use of different medication combinations for the pre-symptomatic survival time of AD onset. Additionally, we employed a supervised machine learning algorithm (random forest) to assess the relative importance of various therapies in predicting the occurrence of AD. Adding Metformin to the combination of ACEI+Beta Blocker (HR = 0.56, 95% CI (0.41, 0.77)) reduced the risk of AD onset compared to ACEI monotherapy alone (HR = 0.91, (0.85, 0.98)), Beta Blocker monotherapy (HR = 0.86, 95% CI (0.80, 0.92)), or combined ACEI+Beta Blocker (HR=0.85, 95%CI (0.77, 0.94)), when statin prescribers were used as a reference. Prescriptions of ARB alone or the combination of ARB with Beta Blocker showed an association with a lower risk of AD onset. Selected medications for the treatment of multiple chronic conditions among elderly individuals with hypertension, hyperlipidemia, and diabetes as monotherapy or combination therapies lengthen the pre-symptomatic period before the onset of AD.


Acute Kidney Injury: Definition, Pathophysiology and Clinical Phenotypes.
Acute kidney injury (AKI) is a clinical syndrome that complicates the course and worsens the outcome in a significant number of hospitalised patients. Recent advances in clinical and basic research will help with a more accurate definition of this syndrome and in the elucidation of its pathogenesis. With this knowledge we will be able to conduct more accurate epidemiologic studies in an effort to gain a better understanding of the impact of this syndrome. AKI is a syndrome that rarely has a sole and distinct pathophysiology. Recent evidence, in both basic science and clinical research, is beginning to change our view for AKI from a single organ failure syndrome to a syndrome where the kidney plays an active role in the progress of multi-organ dysfunction. Accurate and prompt recognition of AKI and better understanding of the pathophysiologic mechanisms underlying the various clinical phenotypes are of great importance to research for effective therapeutic interventions. In this review we provide the most recent updates in the definition, epidemiology and pathophysiology of AKI.


The potential biotechnological applications of the exopolysaccharide produced by the halophilic bacterium Halomonas almeriensis.
We have studied the extracellular polysaccharide (EPS) produced by the type strain, M8(T), of the halophilic bacterium Halomonas almeriensis, to ascertain whether it might have any biotechnological applications. All the cultural parameters tested influenced both bacterial growth and polysaccharide production. EPS production was mainly growth-associated and under optimum environmental and nutritional conditions M8(T) excreted about 1.7 g of EPS per litre of culture medium (about 0.4 g of EPS per gram of dry cell weight). Analysis by anion-exchange chromatography and high-performance size-exclusion chromatography indicated that the exopolysaccharide was composed of two fractions, one of 6.3 × 10(6) and another of 1.5 × 10(4) Daltons. The monosaccharide composition of the high-molecular-weight fraction was mannose (72% w/w), glucose (27.5% w/w) and rhamnose (0.5% w/w). The low-molecular-weight fraction contained mannose (70% w/w) and glucose (30% w/w). The EPS has a substantial protein fraction (1.1% w/w) and was capable of emulsifying several hydrophobic substrates, a capacity presumably related to its protein content. The EPS produced solutions of low viscosity with pseudoplastic behaviour. It also had a high capacity for binding some cations. It contained considerable quantities of sulphates (1.4% w/w), an unusual feature in bacterial polysaccharides. All these characteristics render it potentially useful as a biological agent, bio-detoxifier and emulsifier.


[Activities in the Senegalese National Center of Enterobacteria in 1980, 1981 and 1982].
1,589 strains of Salmonella and 192 strains of Shigella have been studied in the Senegalese National Center of Enterobacteria during 1980, 1981 and 1982. By the side of typhoid fever which remains the most frequent salmonellosis in Senegal (51.4% of human isolates), hospital-acquired salmonellosis have to be separately dealt with ( S. ordonez in 1980, S. chester in 1981, S. ibadan in 1982), characterized by high infantile mortality and constant multiple resistance to antibiotics. A country investigation showed the carriage of numerous and different serotypes in men and animals. Shigella strains isolated in Senegal keep yearly a similar antigenic distribution. On the other hand, occurrence of resistant strains (to ampicillin, tetracycline, trimethoprim -sulfamethoxazole) seems to be increasing.


Addressing a growing community's health needs: project SAHNA (South Asian Health Needs Assessment).
Limited data and research is available on the health issues faced by the South Asian population, especially for those living in the Washington, DC metropolitan area. Online and in-person surveys were administered to a convenience sample (n = 709) of South Asians living in the metropolitan Washington DC region. The survey gathered information on socio-demographics and acculturation; health care access and utilization; sources of health information; perceptions of community health; physical activity and smoking. While over 70% of participants had a routine physical exam in the last 12 months, foreign born and less acculturated adults were less likely to have had a routine visit in the last 12 months. Internet (76.9%) and physicians (76.1%) are key sources of health information for South Asians. Only 29% of South Asians are engaging in the recommended amount of physical activity per week. The results of this study provide guidance for future work in addressing the health and well-being of South Asian communities in the United States.