An automated multidimensional thin film stretching device for the generation of anisotropic polymeric micro- and nanoparticles.
Anisotropic polymeric particles are of growing interest for biomaterials applications due to their unique properties. These include the ability for these particles to evade nonspecific cellular uptake and to have enhanced targeted cellular uptake and interaction. One of the most widely used methods for generating anisotropic polymeric particles is the thin film stretching procedure. Despite its theoretical simplicity, this procedure, as it has been implemented to date, can be difficult due to the inconsistent nature of the manual operation of machinery used to stretch the film. We have constructed an automated thin film stretcher for control over biomaterials via thin film stretching in 1D and 2D and as a result, have enabled precise generation of anisotropic polymeric particles. We demonstrate that this device can be utilized to produce anisotropic biodegradable particles of different size, shape, and material consistency. Furthermore, we show that this machine has enabled the scaled up and rapid production of anisotropic polymeric particles, including polymeric microparticles that mimic the shape of red blood cells. Further application of this automated thin film stretching device could allow for significant impact to diverse biomaterial and biomedical applications such as biomimetic particles for immunoengineering and long-circulating particles for controlled release of drugs.


What does it mean to be an oncology nurse? Reexamining the life cycle concepts.
To summarize the current research pertaining to the concepts initially examined by the Oncology Nursing Society Life Cycle of the Oncology Nurse Task Force and related projects completed in 1994. Published articles on the 21 concepts from the Oncology Nursing Society Life Cycle of the Oncology Nurse Task Force work. Research published in English from 1995-2009 was obtained from PubMed, CINAHL(R), PsycINFO, ISI Science, and EBSCO Health Source(R): Nursing/Academic Edition databases. Most of the concepts identified from the Oncology Nursing Society Life Cycle of the Oncology Nurse Task Force have been examined in the literature. Relationships and witnessing suffering were common concepts among studies of the meaning of oncology nursing. Nurses provide holistic care, and not surprisingly, holistic interventions have been found useful to support nurses. Interventions included storytelling, clinical support of nurses, workshops to find balance in lives, and dream work. Additional support comes from mentoring. The research identified was primarily descriptive, with very few interventions reported. Findings have been consistent over time in diverse countries. This review indicates that although the healthcare system has changed significantly in 15 years, nurses' experiences of providing care to patients with cancer have remained consistent. The need for interventions to support nurses remains.


Causal association of serum biomarkers with oral cavity and oropharyngeal cancer: a mendelian randomization study.
Observational epidemiological studies revealed that multiple serum biomarkers can be associated with the risk of oral and oropharyngeal cancer (OC/OPC). However, the causal relationship between them remains largely unknown. This study aimed to investigate the causal relationship between potential serum biomarkers and (OC/OPC). A two-sample Mendelian randomization (MR) approach was performed to assess the causal association of 10 serum biomarkers with the risk of OC / OPC. Summary data on OC/OPC were obtained from a GWAS meta-analysis that included 2497 cases and 2928 controls. The TwoSampleMR package in R was used to perform MR analyzes. Inverse-variance weighted (IVW), Weighted median and MR-Egger methods were used to assess causal effects. Suggestive associations with increased risk of C-reactive protein (CRP) (OR 1.52, 95% CI 1.14 to 2.02), using the IVW method. MR-Egger regression suggested that directional pleiotropy was unlikely to bias the result (P = 0.19). The findings were robust to sensitivity analyzes. The risk of OC/OPC was not associated with serum 25-hydroxyvitamin D, HDL cholesterol, LDL cholesterol, total cholesterol, triglycerides, adiponectin, leptin, HbA1C and Insulin-like growth factor 1 (IGF 1). This study supports that CRP was causally associated with an increased risk of oral and oropharyngeal cancer.


Cdkn2a is an atherosclerosis modifier locus that regulates monocyte/macrophage proliferation.
Common genetic variants in a 58-kb region of chromosome 9p21, near the CDKN2A/CDKN2B tumor suppressor locus, are strongly associated with coronary artery disease. However, the underlying mechanism of action remains unknown. We previously reported a congenic mouse model harboring an atherosclerosis susceptibility locus and the region of homology with the human 9p21 locus. Microarray and transcript-specific expression analyses showed markedly decreased Cdkn2a expression, including both p16(INK4a) and p19(ARF), but not Cdkn2b (p15(INK4b)), in macrophages derived from congenic mice compared with controls. Atherosclerosis studies in subcongenic strains revealed genetic complexity and narrowed 1 locus to a small interval including Cdkn2a/b. Bone marrow (BM) transplantation studies implicated myeloid lineage cells as the culprit cell type, rather than resident vascular cells. To directly test the role of BM-derived Cdkn2a transcripts in atherogenesis and inflammatory cell proliferation, we performed a transplantation study using Cdkn2a(-/-) cells in the Ldlr(-/-) mouse model. Cdkn2a-deficient BM recipients exhibited accelerated atherosclerosis, increased Ly6C proinflammatory monocytes, and increased monocyte/macrophage proliferation compared with controls. These data provide a plausible mechanism for accelerated atherogenesis in susceptible congenic mice, involving decreased expression of Cdkn2a and increased proliferation of monocyte/macrophages, with possible relevance to the 9p21 human locus.


Clinical evaluation of combined spatial compounding and adaptive imaging in breast tissue.
When spatial compounding is applied to targets with significant acoustic velocity inhomogeneities, the correlation between speckle patterns of the images to be averaged decreases, thereby increasing the speckle reduction nominally obtained. Phase correction applied to these targets improves the coherence of the wavefield and restores image spatial frequencies. Combining these two modes can be used to effectively increase the contrast-to-noise ratio (CNR) of imaging targets and improve the general image quality of these targets over spatial compounding alone. This paper presents a clinical evaluation of combined spatial compounding and adaptive imaging in breast tissue and compares this combined technique to conventional imaging and to adaptive imaging and spatial compounding operating independently. Experiments were performed on a 1.75-D, 8 x 96 array attached to a commercially-available scanner. Cysts, microcalcifications and other breast structures were targeted in order to assess the impact of the combined mode on CNR, target width, target brightness and target peak-to-background ratio (PBR). In general, phase correction improved cyst CNR by 7.7%, decreased target width by 18.7%, increased target brightness by 30.1% and increased PBR by 17.9%. Compounding alone, using three overlapping 9.71 mm subapertures, increased cyst CNR by 24.6%, but increased target width by 25.4% and decreased PBR by 13.2%. Combining both modes, however, increased cyst CNR by 32.6%, inappreciably increased target width by 1.1% and marginally decreased PBR by 2.8%. The increase in target brightness with this combined mode was 20.0%


Shearwave Elastography Increases Diagnostic Accuracy in Characterization of Breast Lesions.
The purpose of this study was to investigate the diagnostic efficacy of shearwave elastography (SWE) in differentiating between benign and malignant breast lesions.One hundred and fifty-nine lesions were assessed using B-mode ultrasound (US) and SWE parameters were recorded (Emax, Emean, Emin, Eratio, SD). SWE measurements were then correlated with histopathological diagnosis.The final sample contained 85 benign and 74 malignant lesions. The maximum stiffness (Emax) with a cutoff point of ≥ 56.0 kPa (based on ROC curves) provided sensitivity of 100.0%, specificity of 97.6%, positive predictive value (PPV) of 97.4%, and negative predictive value (NPV) of 100% in detecting malignant lesions. A cutoff of ≥80 kPa managed to downgrade 95.5% of the Breast Imaging-Reporting and Data System (BI-RADS) 4a lesions to BI-RADS 3, negating the need for biopsy. Using a combination of BI-RADS and SWE, the authors managed to improve the PPV from 2.3% to 50% in BI-RADS 4a lesions.SWE of the breast provides highly specific and sensitive quantitative values that are beneficial in the characterization of breast lesions. Our results showed that Emax is the most accurate value for differentiating benign from malignant lesions.


[Ophthalmoscopic findings in 3 patients with panarteritis nodosa and review of the literature].
Ocular involvement in panarteritis nodosa (PAN) has been reported to occur in 10 to 20% of patients. In 3 patients with acute visual disturbance we point out unusual findings. Case 1. A 40-year-old man initially presented with papilledema together with partial optic atrophy in both eyes, later polyneuropathy, gangrene of the toes and myalgic pains developed. Caliber changes in the small arteries in the liver were seen angiographically and recognized as signs of PAN. Under treatment with cyclophosphamide und prednisone no relapse occurred during a follow-up of 2 years. Case 2. In a 67-year-old man who suffered from arterial hypertension and coronary heart disease, central retinal artery occlusion occurred, at first in the left and then later in the right eye. The clinically suspected diagnosis of PAN (arterial hypertension, myalgia, polyneuropathy) was confirmed by a muscle biopsy. During a follow-up of 4 years--including treatment with prednisone and cyclophosphamide--no relapse occurred. Case 3. A 16-year-old adolescent with throbbing headaches and a thickened right temporal artery reported visual disturbances. These were due to an inflammation of choroidal vessels of the right eye appearing as an initial sign of PAN. Histology revealed a necrotising arteritis of the temporal artery. He presented with signs of Raynaud's disease, cachexia and arterial hypertension. Multiple vasculitic changes were detected by aorto-arteriography. Five months after the visual deterioration an anterior spinal artery syndrome with quadriplegia developed. After a follow-up of 2 years and treatment with prednisone und cyclophosphamide, he still had paralysis of both legs. The visual acuity was 1.0 in each eye. PAN should be considered in differential diagnosis in patients with acute inflammatory signs of the optic nerve head, the choroid and/or the retina together with general signs of the disease. If the disease is suspected, a muscle biopsy is indicated.


NikA/TcsC histidine kinase is involved in conidiation, hyphal morphology, and responses to osmotic stress and antifungal chemicals in Aspergillus fumigatus.
The fungal high osmolarity glycerol (HOG) pathway is composed of a two-component system (TCS) and Hog1-type mitogen-activated protein kinase (MAPK) cascade. A group III (Nik1-type) histidine kinase plays a major role in the HOG pathway of several filamentous fungi. In this study, we characterized a group III histidine kinase, NikA/TcsC, in the life-threatening pathogenic fungus, Aspergillus fumigatus. A deletion mutant of nikA showed low conidia production, abnormal hyphae, marked sensitivity to high osmolarity stresses, and resistance to cell wall perturbing reagents such as congo red and calcofluor white, as well as to fungicides such as fludioxonil, iprodione, and pyrrolnitrin. None of these phenotypes were observed in mutants of the SskA response regulator and SakA MAPK, which were thought to be downstream components of NikA. In contrast, in response to fludioxonil treatment, NikA was implicated in the phosphorylation of SakA MAPK and the transcriptional upregulation of catA, dprA, and dprB, which are regulated under the control of SakA. We then tested the idea that not only NikA, but also the other 13 histidine kinases play certain roles in the regulation of the HOG pathway. Interestingly, the expression of fos1, phkA, phkB, fhk5, and fhk6 increased by osmotic shock or fludioxonil treatment in a SakA-dependent manner. However, deletion mutants of the histidine kinases showed no significant defects in growth under the tested conditions. Collectively, although the signal transduction network related to NikA seems complicated, NikA plays a crucial role in several aspects of A. fumigatus physiology and, to a certain extent, modulates the HOG pathway.


[Surgical treatment of syndactylia in recessive dystrophic epidermolysis bullosa (author's transl)].
The authors report on their experience with the surgical treatment of syndactylia and flexor contracture in recessive dystrophic epidermolysis bullosa in a 7 1/2-year-old boy. After separation of the fused fingers by excision of the epidermal sac, all sclerotic tissue and affected skin, the defects are covered by split autograft skin taken from clinically non-affected areas. Morphologic and functional results are very satisfactory.


Brain representations of negative numbers.
Participants performed a physical comparison task of pairs of positive and pairs of negative one-digit numbers while their electrophysiological brain activity was measured. The numerical value of the presented digits was either congruent or incongruent with the physical size of the digits. Analysis has shown that the earliest event-related potential (ERP) difference between positive and negative numbers was found in the P300 ERP component peak, where there was an inverse effect of congruity in the negative pairs, compared with the positive ones. This pattern of results supports the idea that natural numbers serve as primitives of the human cognitive system, whereas negative numbers are apparently generated if needed.


A feeding protocol for delivery of agents to assess development in Varroa mites.
A novel feeding protocol for delivery of bio-active agents to Varroa mites was developed by providing mites with honey bee larva hemolymph supplemented with cultured insect cells and selected materials delivered on a fibrous cotton substrate. Mites were starved, fed on treated hemolymph to deliver selected agents and then returned to bee larvae. Transcript levels of two reference genes, actin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH), as well as for nine selected genes involved in reproductive processes showed that the starvation and feeding protocol periods did not pose a high level of stress to the mites as transcript levels remained comparable between phoretic mites and those completing the protocol. The feeding protocol was used to deliver molecules such as hormone analogs or plasmids. Mites fed with Tebufenozide, an ecdysone analog, had higher transcript levels of shade than untreated or solvent treated mites. In order to extend this feeding protocol, cultured insect cells were incorporated to a final ratio of 1 part cells and 2 parts hemolymph. Although supplementation with Bombyx mori Bm5 cells increased the amount of hemolymph consumed per mite, there was a significant decrease in the percentage of mites that fed and survived. On the other hand, Drosophila melanogaster S2 cells reduced significantly the percentage of mites that fed and survived as well as the amount of hemolymph consumed. The feeding protocol provides a dynamic platform with which to challenge the Varroa mite to establish efficacy of control agents for this devastating honey bee pest.


American College of Surgeons' Committee on Trauma Performance Improvement and Patient Safety program: maximal impact in a mature trauma center.
To examine the impact of an ongoing comprehensive performance improvement and patient safety (PIPS) program implemented in 2005 on mortality outcomes for trauma patients at an established American College of Surgeons (ACS)-verified Level I Trauma Center. The primary outcome measure was in-hospital mortality. Age, Injury Severity Score (ISS), and intensive care unit admissions were used as stratifying variables to examine outcomes over a 5-year period (2004-2008). Institution mortality rates were compared with the National Trauma Data Bank mortality rates stratified by ISS score. Enhancements to our comprehensive PIPS program included revision of trauma activation criteria, development of standardized protocols for initial resuscitation, massive transfusion, avoidance of over-resuscitation, tourniquet use, pelvic fracture management, emphasis on timely angiographic and surgical intervention, prompt spine clearance, reduction in time to computed tomography imaging, reduced dwell time in emergency department, evidence-based traumatic brain injury management, and multidisciplinary efforts to reduce healthcare-associated infections. In 2004 (baseline data), the in-hospital mortality rate for the most severely injured trauma patients (ISS >24) at our trauma center was 30%, consistent with the reported mortality rate from the National Trauma Data Bank for patients with this severity of injury. Over 5 years, our mortality rate decreased significantly for severely injured patients with an ISS >24, from 30.1% (2004) to 18.3% (2008), representing a 12% absolute reduction in mortality (p = 0.011). During the same 5-year time period, the proportion of elderly patients (age >65 years) cared for at our trauma center increased from 23.5% in 2004 to 30.6% in 2008 (p = 0.0002). Class I trauma activations increased significantly from 5.5% in 2004 to 15.5% in 2008 based on our reclassification. A greater percentage of patients were admitted to the intensive care unit (25.8% in 2004 to 37.3% in 2007 and 30.4% in 2008). No difference was identified in the rate of blunt (95%) or penetrating (5%) mechanism of injury in our patients over this time period. Trauma Quality Improvement Program confirmed improved trauma outcomes with observed-to-expected ratio and 95% confidence intervals of 0.64 (0.42-0.86) for all patients, 0.54 (0.15-0.91) for blunt single-system patients, and 0.78 (0.51-1.06) for blunt multisystem patients. Implementation of a multifaceted trauma PIPS program aimed at improving trauma care significantly reduced in-hospital mortality in a mature ACS Level I trauma center. Optimal care of the injured patient requires uncompromising commitment to PIPS.


Growing healthy families: family use of pedometers to increase physical activity and slow the rate of obesity.
We conducted a study to determine if wearing a pedometer affects weight, body mass index (BMI), or mediators of physical activity among families. Eighty-seven families were randomized to 1 of 3 treatments: pedometer plus education (PE), pedometer (P), or control (C). Participants in the PE and P groups wore pedometers and were encouraged to walk 10,000 steps daily for 12 weeks. PE group participants attended 6 sessions on healthy eating and exercise. Participants were surveyed about their knowledge and attitudes about healthy eating and physical activity prior to randomization, at the end of the intervention, and 9 months later. Their heights and weights were measured and BMI calculated. Children's BMI percentile decreased from baseline to end of intervention (-0.18%) and at 9-month follow-up (-0.08%) but did not differ by treatment. Children's BMI percentile varied by parental obesity status (average BMI percentile was 88.7% for children of obese parents and 78.5% for children of non-obese parents). Parents' weight decreased slightly by intervention's end (0.6 pounds) and at 9 months (1.2 pounds), but change was similar among groups. Attitudes about their physical activity level relative to their peers improved significantly among children and parents wearing the pedometer. Self-efficacy improved for parents wearing the pedometer. Both children and parents felt the pedometer increased their activity level, but most were unlikely to wear it beyond the intervention. The pedometer had little impact on the activity level, weight, or BMI of participants.


Superprotonic conductivity in RbH2-3y(PO4)1-y: a phosphate deficient analog to cubic CsH2PO4 in the (1 - x)RbH2PO4 - xRb2HPO4 system.
In contrast to CsH2PO4 (cesium dihydrogen phosphate, CDP), a material with a well-established superprotonic transition to a high conductivity state at 228 °C, RbH2PO4 (rubidium dihydrogen phosphate, RDP) decomposes upon heating under ambient pressure conditions. Here we find, from study of the (1 - x)RbH2PO4 - xRb2HPO4 system, the remarkable occurrence of cubic, off-stoichiometric RbH2-3y(PO4)1-y, or α-RDP, with a variable Rb : PO4 ratio. Materials were characterized by simultaneous thermal analysis and in situ X-ray powder diffraction performed under high steam partial pressure, from which the phase diagram between RbH2PO4 (x = 0) and Rb5H7(PO4)4 (x = 1/4) was established. The system displays eutectoid behavior, with a eutectoid transition temperature of 242.0 ± 0.5 °C and eutectoid composition of x = 0.190 ± 0.004. Even the end-member Rb5H7(PO4)4 appears to transform to α-RDP, implying y in the chemical formula of 0.2 and a phosphate site vacancy concentration as high as 20%. Charge balance is attained by a decrease in the average number of protons on the remaining phosphate groups. The cubic lattice parameter at x = 0.180, near the eutectoid composition, and at a temperature of 249 °C is 4.7138(2) Å. This value is substantially smaller than the estimated ambient-pressure lattice parameter of stoichiometric RbH2PO4 of 4.837(12) Å, consistent with the proposal of phosphate site vacancies in the former. The superprotonic conductivity of the x = 0.180 material is 6 × 10-3 S cm-1 at 244 °C, a factor of three lower than that of CDP at the same temperature. While the engineering properties of α-RDP do not suggest immediate technological relevance, the discovery of a superprotonic solid acid with a high concentration of phosphate site vacancies opens new avenues for developing proton conducting electrolytes, and in particular, for controlling their transition behavior.


Dynamic depth-dependent osmotic swelling and solute diffusion in articular cartilage monitored using real-time ultrasound.
The objective of this study was to investigate the feasibility of ultrasonic monitoring for the transient depth-dependent osmotic swelling and solute diffusion in normal and degenerated articular cartilage (artC) tissues. Full-thickness artC specimens were collected from fresh bovine patellae. The artC specimens were continuously monitored using a focused beam of 50 MHz ultrasound (US) during sequential changes of the bathing solution from 0.15 mol/L to 2 mol/L saline, 0.15 mol/L saline, 1 mg/mL trypsin solution, 0.15 mol/L saline, 2 mol/L saline and back to 0.15 mol/L saline. The transient displacements of US echoes from the artC tissues at different depths were used to represent the tissue deformation and the NaCl diffusion. The trypsin solution was used selectively to digest the proteoglycans in artC. It was demonstrated that high-frequency US was feasible for monitoring the transient osmotic swelling, solute transport and progressive degeneration of artC in real-time. Preliminary results showed that the normal bovine patellar artC shrank during the first several minutes and then recovered to its original state in approximately 1 h when the solution was changed from 0.15 mol/L to 2 mol/L saline. Degenerated artC showed neither shrinkage nor recovery during the same process. In addition, a dehydrated-hydrated artC specimen showed much stronger shrinkage and it resumed the original state when the solution was changed from 2 mol/L back to 0.15 mol/L saline. The diffusion of NaCl and the digestion process of proteoglycans induced by trypsin were also successfully monitored in real-time.


Canine hypothyroidism: a review of aetiology and diagnosis.
Hypothyroidism is recognised as an important endocrine disorder of dogs, and a frequent differential for numerous presenting complaints. Its diagnosis has never been straight forward as results suggestive of hypothyroidism can occur for a variety of reasons in dogs with normal thyroid function (euthyroid). As a consequence, the accurate investigation of hypothyroidism has been hindered by the potential inclusion of a number of cases not truly hypothyroid. In recent years, the development of newer diagnostic tests, e.g. free thyroxine, canine thyroid stimulating hormone, thyroglobulin autoantibodies, has significantly improved our ability to reliably differentiate hypothyroidism from other clinically similar disorders. This has led to a marked increase in our knowledge of the phenotypic, genotypic and aetiological aspects of this disorder in dogs.


Wt1-expressing progenitors contribute to multiple tissues in the developing lung.
Lungs develop from paired endodermal outgrowths surrounded by a mesodermal mesenchyme. Part of this mesenchyme arises from epithelial-mesenchymal transition of the mesothelium that lines the pulmonary buds. Previous studies have shown that this mesothelium-derived mesenchyme contributes to the smooth muscle of the pulmonary vessels, but its significance for lung morphogenesis and its developmental fate are still little known. We have studied this issue using the transgenic mouse model mWt1/IRES/GFP-Cre (Wt1cre) crossed with the Rosa26R-EYFP reporter mouse. In the developing lungs, Wt1, the Wilms' tumor suppressor gene, is specifically expressed in the embryonic mesothelium. In the embryos obtained from the crossbreeding, the Wt1-expressing cell lineage produces the yellow fluorescent protein (YFP), allowing for colocalization with differentiation markers. Wt1cre-YFP cells were very abundant from the origin of the lung buds to postnatal stages, contributing significantly to pulmonary endothelial and smooth muscle cells, bronchial musculature, tracheal and bronchial cartilage, as well as CD34⁺ fibroblast-like interstitial cells. Thus Wt1cre-YFP mesenchymal cells show the very same differentiation potential as the splanchnopleural mesenchyme surrounding the lung buds. FSP1⁺ fibroblast-like cells were always YFP⁻; they expressed the common leukocyte antigen CD45 and were apparently recruited from circulating progenitors. We have also found defects in pulmonary development in Wt1-/- embryos, which showed abnormally fused lung lobes, round-shaped and reduced pleural cavities, and diaphragmatic hernia. Our results suggest a novel role for the embryonic mesothelium-derived cells in lung morphogenesis and involve the Wilms' tumor suppressor gene in the development of this organ.


Secretory IgA antibodies from plants.
Secretory IgA (SIgA) is the antibody type produced in both mammals and birds that protects the body from infection at mucosal surfaces. While monoclonal IgG antibodies, particularly those against tumor antigens, have received a great deal of attention, both scientific and commercial, as immunotherapeutic agents, the potential of SIgA antibodies has only recently begun to be exploited. Part of the reason for this is that SIgA production in vivo normally requires the cooperation of two different cell types, and single animal cell systems for monoclonal SIgA production are inefficient. Transgenic plants are currently the most productive and economical system for making SIgA. The only monoclonal SIgA to be tested therapeutically in a human clinical trial is a product called CaroRx, made in transgenic tobacco, which is designed to block adherence to teeth of the bacteria that causes cavities. This antibody accumulates to high levels in the leaves of tobacco, where it is located primarily in the endoplasmic reticulum. The antibody can be efficiently purified using the affinity reagent protein G. Topical oral treatment in human subjects was safe and effective. Characterization of the expression, secretion, purification and therapeutic use of this antibody serves as a model for additional plant-made therapeutic SIgA antibodies under development.


Effect of the inoculation site of bovine purified protein derivative (PPD) on the skin fold thickness increase in cattle from officially tuberculosis free and tuberculosis-infected herds.
The official technique for diagnosis of bovine tuberculosis (bTB) worldwide is the tuberculin skin test, based on the evaluation of the skin thickness increase after the intradermal inoculation of a purified protein derivative (PPD) in cattle. A number of studies performed on experimentally infected or sensitized cattle have suggested that the relative sensitivity of the cervical test (performed in the neck) may vary depending on the exact location in which the PPD is injected. However, quantitative evidence on the variation of the test accuracy associated to changes in the site of inoculation in naturally infected animals (the population in which performance of the test is most critical for disease eradication) is lacking. Here, the probability of obtaining a positive reaction (>2 or 4 millimeters and/or presence of local clinical signs) after multiple inoculations of bovine PPD in different cervical and scapular locations was assessed in animals from five bTB-infected herds (818 cattle receiving eight inoculations) using a hierarchical Bayesian logistic regression model and adjusting for the potential effect of age and sex. The effect of the inoculation site was also assessed qualitatively in animals from four officially tuberculosis free (OTF) herds (two inoculations in 210 animals and eight inoculations in 38 cattle). Although no differences in the qualitative outcome of the test were observed in cattle from OTF herds, a statistically important association between the test outcome and the inoculation site in animals from infected herds was observed, with higher probabilities of positive results when the test was performed in the neck anterior area. Our results suggest that test sensitivity may be maximized by considering the area of the neck in which the test is applied, although lack of effect of the inoculation site in the specificity of the test should be confirmed in a larger sample.


Evidence for direct linkage between antigen recognition and lytic expression in effector T cells.
The relationship between antigen recognition and lytic expression by effector T cells was examined by coculturing two cytolytically active lymphoid cell populations. When antigen recognition between the populations could occur only in one direction, then cytotoxicity was expressed only in that direction and the population whose antigens were recognized lost its lytic activity. In contrast, the cocultured effector cell population fully maintained its lytic potential. This lack of reciprocal inactivation was taken as evidence that T-cell receptor accomodation by surface antigen is linked to the expression of cytolytic activity by effector T lymphocytes.


Interpretation of clinical trials in diffuse large-cell lymphoma.
Diffuse large-cell lymphoma is one of the neoplasms curable with chemotherapy in an appreciable percentage of patients. However, all patients are not cured and the best combination of agents is not certain. This reflects the lack of completed comparative trials using the regimens that appear most effective. Despite this uncertainty, several principles for the therapy of diffuse large-cell lymphoma can be identified that allow an analysis of the results reported in the literature. These principles include the following: (1) for a regimen to be curative in a substantial number of patients it must achieve a high rate of complete remissions; (2) cure must be accomplished with frontline therapy; (3) drugs must be delivered at curative doses; (4) rapidity of achieving a complete response might be related to chance for cure; (5) prolonged treatment for diffuse large-cell lymphoma is unnecessary; and (6) aggressive therapy is toxic. In analyzing the results with any regimen it is important to have long follow-up since late relapses do occur and initial very positive results tend to decay with greater numbers of patients treated. Applying these principles to the reported chemotherapy studies in patients with diffuse large-cell lymphoma suggest that no one of the new regimens is clearly superior to the others. Also, it is not clear that cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) delivered at full doses to comparable patients is inferior to the newer regimens. The results of ongoing studies comparing these regimens might help resolve these questions.


[Effect of electroacupuncture on ethology and cytokines of hippocampus in rats with dysmnesy].
To observe the effect of electroacupuncture (EA) on learning and memory impairment and cytokines of hippocampus in aging rats induced by D-galactose for exploring its underlying mechanism in the treatment of dysmnesy. A total of 27 SD rats were randomly divided into control group (n = 9), model group (n = 8) and EA group (n = 10). Dysmnesy model was induced by intraperitoneal injection of D-galactose. EA (3 Hz, continuous waves, 1 mA) was applied to "Baihui" (GV 20) and "Zusanli" (ST 36) for 20 min every time and on alternate days, continuously for 21 days. Morris water maze tests were conducted to detect the rat's escape latency, percentage of swimming distance in the original platform quadrant and the total distance (SD/TD) and the percentage of swimming time in the platform quadrant after removal of the platform (spatial probe test) which were used for assessing the animals' learning and memory ability. The contents of IL-1beta, IL-6 and TNF-alpha in the hippocampus tissue were assayed by radioimmunoassay. Location navigation test showed that the escape latencies on the 2nd day and the 3rd day in model group were significantly longer than those of control group (P < 0.05), and the percents of SD/TD and swimming time in the platform quadrant were significantly lower than those of control group (P < 0.05, P < 0.01). In comparison with model group, the escape latency of EA group on the 3rd day was significantly shorter (P < 0.05), and the percent of SD/TD in the platform quadrant of EA group was significantly longer (P < 0.01). Compared with control group, IL-1beta and TNF-alpha contents in the hippocampus increased significantly in model group (P < 0.05, P < 0.01), while IL-6 content in model group decreased significantly (P < 0.01). In comparison with model group, the levels of IL-1beta and TNF-alpha in EA group decreased significantly (P < 0.05). EA can improve the learning-memory ability in dysmnesy rats, which may be closely associated with its effects in regulating the levels of cytokines in the hippocampus.


Transdermal penetration behaviour of drugs: CART-clustering, QSPR and selection of model compounds.
A set of 116 structurally very diverse compounds, mainly drugs, was characterized by 1630 molecular descriptors. The biological property modelled in this study was the transdermal permeability coefficient logK(p). The main objective was to find a limited set of suitable model compounds for skin penetration studies. The classification and regression trees (CART) approach was applied and the resulting groups were discussed in terms of their role as possible model compounds and their determining descriptors. A second objective was to model transdermal penetration as a function of selected descriptors in quantitative structure-property relationships (QSPR) using a boosted CART (BRT) approach and multiple linear regression (MLR) analysis, where regression models were obtained by stepwise selection of the best descriptors. Evaluation of the standard statistical, as well as descriptor-number dependent, regression quality attributes yielded a maximal 10-dimensional MLR model. The CART and MLR models were subjected to an external validation with a test set of 12 compounds, not included in the original learning set of 104 compounds, to assess the predictive power of the models.


[The use of topiramate in the treatment of focal epilepsy in children].
Topiramate was used in the treatment of 66 children, aged 2--16 years (mean age 7.0±4.6 years), including 19 patients with idiopathic form of focal epilepsy (IFE), 21 patients with cryptogenic forms (CFE) and 26 patients with symptomatic forms (SFE). The drug was administered in capsules in dose from 3 to 7 mg/kg/day. Thirty-three patients were on monotherapy, 26 patients received the drug in the combination with one antiepileptic (AED) drug and 7 patients received topiramate in the combination with 2 AEDs. The efficacy of topiramate in focal epilepsy was demonstrated in 65% of patients, including high efficacy in 15% and complete reduction of seizures in 12%. No effect was seen in 27% of patients. The results revealed the high efficacy of topiramate in all subgroups of patients. The significantly higher efficacy was noted in patients with IFE and CFE forms compared to those with SFE form. The tolerability was satisfactory in all subgroups. Side-effects observed in 5 patients were eliminated by increasing the duration of dose titration.


The dual action of glioma-derived exosomes on neuronal activity: synchronization and disruption of synchrony.
Seizures represent a frequent symptom in gliomas and significantly impact patient morbidity and quality of life. Although the pathogenesis of tumor-related seizures is not fully understood, accumulating evidence indicates a key role of the peritumoral microenvironment. Brain cancer cells interact with neurons by forming synapses with them and by releasing exosomes, cytokines, and other small molecules. Strong interactions among neurons often lead to the synchronization of their activity. In this paper, we used an in vitro model to investigate the role of exosomes released by glioma cell lines and by patient-derived glioma stem cells (GSCs). The addition of exosomes released by U87 glioma cells to neuronal cultures at day in vitro (DIV) 4, when neurons are not yet synchronous, induces synchronization. At DIV 7-12 neurons become highly synchronous, and the addition of the same exosomes disrupts synchrony. By combining Ca2+ imaging, electrical recordings from single neurons with patch-clamp electrodes, substrate-integrated microelectrode arrays, and immunohistochemistry, we show that synchronization and de-synchronization are caused by the combined effect of (i) the formation of new neuronal branches, associated with a higher expression of Arp3, (ii) the modification of synaptic efficiency, and (iii) a direct action of exosomes on the electrical properties of neurons, more evident at DIV 7-12 when the threshold for spike initiation is significantly reduced. At DIV 7-12 exosomes also selectively boost glutamatergic signaling by increasing the number of excitatory synapses. Remarkably, de-synchronization was also observed with exosomes released by glioma-associated stem cells (GASCs) from patients with low-grade glioma but not from patients with high-grade glioma, where a more variable outcome was observed. These results show that exosomes released from glioma modify the electrical properties of neuronal networks and that de-synchronization caused by exosomes from low-grade glioma can contribute to the neurological pathologies of patients with brain cancers.


Inheritance and the pigmentary dispersion syndrome.
The authors studied 33 patients in five families and have found that 14 of these patients fulfill criteria for the pigmentary dispersion syndrome. Affected patients had characteristics associated with the pigmentary dispersion syndrome, which are: peripheral slit-like iris transillumination defects, increased trabecular meshwork pigmentation, Krukenberg spindle, myopia, and elevated intraocular pressure. The authors observed this syndrome to be transmitted in a direct linear manner from parent to sibling in three of the four families. The transmission of this syndrome was found to be independent of refractive error, iris color, and sexual predilection, all of which are presented.


Large scale modulation of high frequency acoustic waves in periodic porous media.
This paper deals with the description of the modulation at large scale of high frequency acoustic waves in gas saturated periodic porous media. High frequencies mean local dynamics at the pore scale and therefore absence of scale separation in the usual sense of homogenization. However, although the pressure is spatially varying in the pores (according to periodic eigenmodes), the mode amplitude can present a large scale modulation, thereby introducing another type of scale separation to which the asymptotic multi-scale procedure applies. The approach is first presented on a periodic network of inter-connected Helmholtz resonators. The equations governing the modulations carried by periodic eigenmodes, at frequencies close to their eigenfrequency, are derived. The number of cells on which the carrying periodic mode is defined is therefore a parameter of the modeling. In a second part, the asymptotic approach is developed for periodic porous media saturated by a perfect gas. Using the "multicells" periodic condition, one obtains the family of equations governing the amplitude modulation at large scale of high frequency waves. The significant difference between modulations of simple and multiple mode are evidenced and discussed. The features of the modulation (anisotropy, width of frequency band) are also analyzed.


Chondroitin sulfate disaccharides modified the structure and function of the murine gut microbiome under healthy and stressed conditions.
Chondroitin sulfate (CS) has been widely used for medical and nutraceutical purposes due to its roles in maintaining tissue structural integrity. We investigated if CS disaccharides may act as a bioactive compound and modulate gut microbial composition in mice. Our data show that CS disaccharides supplementation for 16 days significantly reduced blood LPS in the mice experiencing exhaustive exercise stress. CS disaccharides partially restored total fecal short-chain fatty acids from the level significantly repressed in mice under the stress. Our findings demonstrated that CS was likely butyrogenic and resulted in a significant increase in fecal butyrate concentration. CS disaccharides had a profound impact on gut microbial composition, affecting the abundance of 13.6% and 7.3% Operational Taxonomic Units in fecal microbial communities in healthy and stressed mice, respectively. CS disaccharides reduced the prevalence of inflammatory Proteobacteria. Together, our findings demonstrated that CS may ameliorate stress-induced intestinal inflammation. Furthermore, CS significantly increased intestinal Bacteroides acidifaciens population, indirectly exerting its immunomodulatory effect on the intestine. CS disaccharides had a significant impact on a broad range of biological pathways under stressed condition, such as ABC transporters, two-component systems, and carbohydrate metabolism. Our results will facilitate the development of CS as a bioactive nutraceutical.


Biolistic transfection and expression analysis of acute cortical slices.
Biolistic gene gun transfection has been used to transfect organotypic cultures (OTCs) or dissociated cultures in vitro. Here, we modified this technique to allow successful transfection of acute brain slices, followed by measurement of neuronal activity within a few hours. We established biolistic transfection of murine acute cortical slices to measure calcium signals. Acute slices are mounted on plasma/thrombin coagulate and transfected with a calcium sensor. Imaging can be performed within 4 h post transfection without affecting cell viability. Four hours after GCaMP6s transfection, acute slices display remarkable fluorescent protein expression level allowing to study spontaneous activity and receptor pharmacology. While optimal gas pressure (150 psi) and gold particle size used (1 μm) confirm previously published protocols, the amount of 5 μg DNA was found to be optimal for particle coating. The major advantage of this technique is the rapid disposition of acute slices for calcium imaging. No transgenic GECI expressing animals or OTC for long periods are required. In acute slices, network interaction and connectivity are preserved. The method allows to obtain physiological readouts within 4 h, before functional tissue modifications might come into effect. Limitations of this technique are random transfection, low expression efficiency when using specific promotors, and preclusion or genetic manipulations that require a prolonged time before physiological changes become measurable, such as expression of recombinant proteins that require transport to distant subcellular localizations. The method is optimal for short-time investigation of calcium signals in acute slices.


Transhiatal (blunt) esophagectomy for malignant and benign esophageal disease: clinical experience and technique.
"Blunt" transhiatal esophagectomy was performed in 23 selected patients. Nineteen had squamous carcinoma of the esophagus (upper third, 1; middle third, 12; distal third, 6), and 2 had adenocarcinoma of the distal esophagus. The other 2 patients had severe lye strictures. Resection with reconstruction was performed in one stage. Esophagogastric continuity was restored using the stomach in the posterior mediastinal position in 20 patients and in the substernal position in 2. The colon in the posterior mediastinal position was used in 1 patient with a lye stricture. Transmural tumor extension or cervical or celiac nodal metastases or both were present in 18 of 21 patients with carcinoma. There was 1 hospital death due to pericardial tamponade. Morbidity included a transient cervical anastomotic leak in 3 patients, one temporary and three permanent unilateral recurrent laryngeal nerve palsies, one intraoperative splenic injury, and severe hemorrhage requiring sternotomy for control in 1 patient. Pulmonary complications occurred in 4 patients: aspiration pneumonia (1) and moderate atelectasis (3). Three patients have died (11, 12, and 17 months postoperatively) in the group with cancer, with follow-up time of 3 to 30 months (mean, 15 months). Transhiatal blunt esophagectomy is a safe and effective procedure in many patients with either esophageal cancer or extensive, benign esophageal strictures.


Antibody-Mediated Inhibition of Tspan12 Ameliorates Vasoproliferative Retinopathy Through Suppression of β-Catenin Signaling.
Anti-angiogenic biologicals represent an important concept for the treatment of vasoproliferative diseases. However, the need for continued treatment, the presence of nonresponders, and the risk of long-term side effects limit the success of existing therapeutic agents. Although Tspan12 has been shown to regulate retinal vascular development, nothing is known about its involvement in neovascular disease and its potential as a novel therapeutic target for the treatment of vasoproliferative diseases. Rodent models of retinal neovascular disease, including the mouse model of oxygen-induced retinopathy and the very low density lipoprotein receptor knockout mouse model were analyzed for Tspan/β-catenin regulation. Screening of a phage display of a human combinatorial antibody (Ab) library was used for the development of a high-affinity Ab against Tspan12. Therapeutic effects of the newly developed Ab on vascular endothelial cells were tested in vitro and in vivo in the oxygen-induced retinopathy and very low density lipoprotein receptor knockout mouse model. The newly developed anti-Tspan12 Ab exhibited potent inhibitory effects on endothelial cell migration and tube formation. Mechanistic studies confirmed that the Ab inhibited the interaction between Tspan12 and Frizzled-4 and effectively modulates β-catenin levels and target genes in vascular endothelial cells. Tspan12/β-catenin signaling was activated in response to acute and chronic stress in the oxygen-induced retinopathy and very low density lipoprotein receptor mouse model of proliferative retinopathy. Intravitreal application of the Ab showed significant therapeutic effects in both models without inducing negative side effects on retina function. Moreover, combined intravitreal injection of the Ab with a known vascular endothelial growth factor inhibitor, Aflibercept, resulted in significant enhancement of the therapeutic efficacy of each monotherapy. Combination therapy with the Tspan12 blocking antibody can be used to reduce anti-vascular endothelial growth factor doses, thus decreasing the risk of long-term off-target effects. Tspan12/β-catenin signaling is critical for the progression of vasoproliferative disease. The newly developed anti-Tspan12 antibody has therapeutic effects in vasoproliferative retinopathy and can enhance the potency of existing anti- vascular endothelial growth factor agents.


Frequency and spectrum of Wolcott-Rallison syndrome in Saudi Arabia: a systematic review.
Wolcott-Rallison syndrome (WRS) is caused by recessive EIF2AK3 gene mutations and characterized by permanent neonatal diabetes (PNDM), skeletal dysplasia, and recurrent hepatitis. The frequency of this rare syndrome is largely unknown. To define the frequency and spectrum of WRS in the Kingdom of Saudi Arabia (KSA) based on published data. The Medline database was searched for published articles on WRS. The number of reported cases from KSA was compared to the total number of WRS cases reported worldwide. The genotype and phenotype of WRS patients from KSA were reviewed. Ten articles describing 23 WRS patients from 12 Saudi families from 1995 to 2012 were identified. This figure accounts for 27.7% (23/83) of the patients and 22.2% (12/54) of the families with WRS reported worldwide until January 2013. All Saudi patients with WRS presented with PNDM, and they represent 59% of all PNDM cases from WRS. At reporting, 73% of patients experienced recurrent hepatitis, 56.5% had skeletal abnormalities, and 39.1% of them were dead. There was a variation in the phenotype even between affected siblings. Genetic diagnosis was confirmed in all 12 families with no correlation between the genotype and phenotype. Eight of the nine EIF2AK3 mutations were only reported in these families, and one was shared with a patient from Qatar, a neighboring Arab state. No study on the frequency of WRS has been published. However, the available data indicate that KSA has the largest collection of patients with WRS worldwide, and nine of the identifiable EIF2AK3 mutations appear to be confined to Arabs. Establishing a national or international registry for WRS would provide more reliable data on this rare condition.


High-sensitivity multi-channel single-interferometer DPSK receiver.
A high-sensitivity method of demodulating wavelength-division-multiplexed optical DPSK channels using a single interferometer is described and demonstrated. We show that this technique can operate within existing standards and achieve near-quantum-limited receiver performance. The resulting hardware simplification has potential to reduce the cost of deploying and scaling future wide-band optical-communication networks employing WDM-DPSK.


The Fabrication and Operation of a Continuous Flow, Micro-Electroporation System with Permeabilization Detection.
Current therapeutic innovations, such as CAR-T cell therapy, are heavily reliant on viral-mediated gene delivery. Although efficient, this technique is accompanied by high manufacturing costs, which has brought about an interest in using alternative methods for gene delivery. Electroporation is an electro-physical, non-viral approach for the intracellular delivery of genes and other exogenous materials. Upon the application of an electric field, the cell membrane temporarily allows molecular delivery into the cell. Typically, electroporation is performed on the macroscale to process large numbers of cells. However, this approach requires extensive empirical protocol development, which is costly when working with primary and difficult-to-transfect cell types. Lengthy protocol development, coupled with the requirement of large voltages to achieve sufficient electric-field strengths to permeabilize the cells, has led to the development of micro-scale electroporation devices. These micro-electroporation devices are manufactured using common microfabrication techniques and allow for greater experimental control with the potential to maintain high throughput capabilities. This work builds off a microfluidic-electroporation technology capable of detecting the level of cell membrane permeabilization at a single-cell level under continuous flow. However, this technology was limited to 4 cells processed per second, and thus a new approach for increasing the system throughput is proposed and presented here. This new technique, denoted as cell-population-based feedback control, considers the cell permeabilization response to a variety of electroporation pulsing conditions and determines the best-suited electroporation pulse conditions for the cell type under test. A higher-throughput mode is then used, where this 'optimal' pulse is applied to the cell suspension in transit. The steps for fabricating the device, setting up and running the microfluidic experiments, and analyzing the results are presented in detail. Finally, this micro-electroporation technology is demonstrated by delivering a DNA plasmid encoding for green fluorescent protein (GFP) into HEK293 cells.


Noninvasive assessment of electroconvulsive-induced changes in cardiac function.
Systolic time intervals were used to noninvasively evaluate the cardiac effects of electroconvulsive therapy (ECT) in eight psychiatric patients. ECT produced a hyperdynamic cardiac state in which there was a significant increase in blood pressure, a shortening of the pre-ejection period (PEP), and an increase in rate pressure product (RPP). Changes in the PEP and the RPP are compatible with increased cardiac contractility and oxygen consumption, respectively. These effects were apparently mediated by an ECT-induced increase in circulating catecholamines, particularly epinephrine.


Very long-term survival and late sudden cardiac death in cardiac resynchronization therapy patients.
The very long-term outcome of patients who survive the first few years after receiving cardiac resynchronization therapy (CRT) has not been well described thus far. We aimed to provide long-term outcomes, especially with regard to the occurrence of sudden cardiac death (SCD), in CRT patients without (CRT-P) and with defibrillator (CRT-D). A total of 1775 patients, with ischaemic or non-ischaemic dilated cardiomyopathy, who were alive 5 years after CRT implantation, were enrolled in this multicentre European observational cohort study. Overall long-term mortality rates and specific causes of death were assessed, with a focus on late SCD. Over a mean follow-up of 30 months (interquartile range 10-42 months) beyond the first 5 years, we observed 473 deaths. The annual age-standardized mortality rates of CRT-D and CRT-P patients were 40.4 [95% confidence interval (CI) 35.3-45.5] and 97.2 (95% CI 85.5-109.9) per 1000 patient-years, respectively. The adjusted hazard ratio (HR) for all-cause mortality was 0.99 (95% CI 0.79-1.22). Twenty-nine patients in total died of late SCD (14 with CRT-P, 15 with CRT-D), corresponding to 6.1% of all causes of death in both device groups. Specific annual SCD rates were 8.5 and 5.8 per 1000 patient-years in CRT-P and CRT-D patients, respectively, with no significant difference between groups (adjusted HR 1.0, 95% CI 0.45-2.44). Death due to progressive heart failure represented the principal cause of death (42.8% in CRT-P patients and 52.6% among CRT-D recipients), whereas approximately one-third of deaths in both device groups were due to non-cardiovascular death. In this first description of very long-term outcomes among CRT recipients, progressive heart failure death still represented the most frequent cause of death in patients surviving the first 5 years after CRT implant. In contrast, SCD represents a very low proportion of late mortality irrespective of the presence of a defibrillator.


Integrated Assessment of the Leading Paths to Mitigate CO2 Emissions from the Organic Chemical and Plastics Industry.
The chemical industry is a major and growing source of CO2 emissions. Here, we extend the principal U.S.-based integrated assessment model, GCAM, to include a representation of steam cracking, the dominant process in the organic chemical industry today, and a suite of emerging decarbonization strategies, including catalytic cracking, lower-carbon process heat, and feedstock switching. We find that emerging catalytic production technologies only have a small impact on midcentury emissions mitigation. In contrast, process heat generation could achieve strong mitigation, reducing associated CO2 emissions by ∼76% by 2050. Process heat generation is diversified to include carbon capture and storage (CCS), hydrogen, and electrification. A sensitivity analysis reveals that our results for future net CO2 emissions are most sensitive to the amount of CCS deployed globally. The system as defined cannot reach net-zero emissions if the share of incineration increases as projected without coupling incineration with CCS. Less organic chemicals are produced in a net-zero CO2 future than those in a no-policy scenario. Mitigation of feedstock emissions relies heavily on biogenic carbon used as an alternative feedstock and waste treatment of plastics. The only scenario that delivers net-negative CO2 emissions from the organic chemical sector (by 2070) combines greater use of biogenic feedstocks with a continued reliance on landfilling of waste plastic, versus recycling or incineration, which has trade-offs.


Peptide-Drug Conjugate Targeting Keratin 1 Inhibits Triple-Negative Breast Cancer in Mice.
Selective delivery of chemotherapy to the tumor site while sparing healthy cells and tissues is an attractive approach for cancer treatment. Carriers such as peptides can facilitate selective tumor targeting and payload delivery. Peptides with specific affinity for the overexpressed cell-surface receptors in cancer cells are conjugated to chemotherapy to afford peptide-drug conjugates (PDCs) that show selective uptake by cancer cells. Using a 10-mer linear peptide (WxEAAYQrFL) called 18-4 that targets and binds breast cancer cells, we designed a peptide 18-4-doxorubicin (Dox) conjugate with high specific toxicity toward triple-negative breast cancer (TNBC) MDA-MB-231 cells and 30-fold lower toxicity to normal breast MCF10A epithelial cells. Here, we elucidate the in vivo activity of this potent and tumor-selective peptide 18-4-Dox conjugate in mice bearing orthotopic MDA-MB-231 tumors. Mice treated with four weekly injections of the conjugate showed significantly lower tumor volumes compared to mice treated with free Dox at an equivalent Dox dose. Immunohistochemical (IHC) analysis of mice tissues revealed that treatment with a low dose of PDC (2.5 mg/kg of Dox equiv) reduced the expression of proliferation markers (PCNA and Ki-67) and increased apoptosis (evidenced by increased caspase-3 expression). At the same dose of free Dox (2.5 mg/kg), the expression of these markers was similar to that of saline treatment. Accordingly, significantly more Dox accumulated in tumors of conjugate-treated mice (7-fold) compared to the Dox-treated mice, while lower levels of Dox were observed in the liver, heart, and lungs of peptide-Dox conjugate-treated mice (up to 3-fold less) than Dox-treated mice. The IHC analysis of keratin 1 (K1), the receptor for peptide 18-4, revealed K1 upregulation in tumors and low levels in normal mammary fat pad and liver tissues from mice, suggesting preferential uptake of PDCs by TNBC to be K1 receptor-mediated. Taken together, our data support the use of a PDC approach to deliver chemotherapy selectively to the TNBC to inhibit tumor growth.


Evaluation of diagnostic and prognostic usefulness of abdominal ultrasonography in dogs with clinical signs of acute pancreatitis.
To report abdominal ultrasonography (AUS) findings in dogs with clinical signs of acute pancreatitis (AP) during the first 2 days of hospitalization and to compare AUS findings with severity of disease and mortality rate. 37 client-owned dogs with clinical signs of AP. Dogs suspected of having AP with complete medical records, AUS examinations performed throughout the first 2 days of hospitalization, and available frozen surplus serum samples for quantitative measurement of canine pancreatic lipase (cPL) concentrations at hospital admission met the criteria for study inclusion. Dogs were grouped as AUS+ or AUS- on the basis of positive or negative findings for AP on AUS, respectively. Abdominal ultrasonography findings of AP were stratified (as mild, moderate, or severe) by use of an AUS severity index, and a canine acute pancreatitis severity score was calculated. 24 of 37 (64.8%) dogs had AUS findings of AP at hospital admission, whereas 10 had positive findings for AP on AUS within 2 days of hospitalization. Three (8%) dogs were AUS- but had serum cPL concentrations > 400 μg/L (ie, values considered diagnostic for AP). On the AUS severity index, 5 of 34 (14.7%) AUS+ dogs had mild findings, 18 (52.9%) AUS+ dogs had moderate findings, and 11 (32.4%) AUS+ dogs had severe findings. Severe findings were associated with a higher risk of death than mild and moderate findings. A significant association was found between canine acute pancreatitis severity scores and mortality rates. For dogs with clinical signs of AP, repeated AUS examinations during hospitalization should be performed, severe findings on the AUS severity index may indicate an increased risk of death, and serum cPL concentrations may increase earlier than findings on AUS of AP.


Major consequences of minor damage: impacts of small grazers on fast-growing kelps.
Damage by small herbivores can have disproportionately large effects on the fitness of individual plants if damage is concentrated on valuable tissues or on select individuals within a population. In marine systems, the impact of tissue loss on the growth rates of habitat-forming algae is poorly understood. We quantified the grazing damage by an isopod Amphoroidea typa on two species of large kelps, Lessonia spicata and Macrocystis pyrifera, in temperate Chile to test whether non-lethal grazing damage could reduce kelp growth rates and photosynthetic efficiency. For L. spicata, grazing damage was widespread in the field, unevenly distributed on several spatial scales (among individuals and among tissue types) and negatively correlated with blade growth rates. In field experiments, feeding by A. typa reduced the concentration of photosynthetic pigments and led to large reductions (~80%) in blade growth rates despite limited loss of kelp biomass (0.5% per day). For M. pyrifera, rates of damage in the field were lower and high densities of grazers were unable to reduce growth rates in field experiments. These results demonstrate that even low per capita grazing rates can result in large reductions in the growth of a kelp, due the spatial clustering of herbivores in the field and the selective removal of photosynthetically active tissues. The impacts of small herbivores on plant performance are thus not easily predicted from consumption rates or abundance in the field, and vary with plant species due to variation in their ability to compensate for damage.


[Prognostic evaluations of the medical and demographic processes].
The medical-and-demographic processes as a starting point for the planning of means and resources for the short- and average-term future are forecasted in the paper on the basis of long-term peculiarities of the natural-science data and with respect for the social-and-economic crisis now underway in the country.


Single-Cell Transcriptome Profiling Unravels Distinct Peripheral Blood Immune Cell Signatures of RRMS and MOG Antibody-Associated Disease.
Relapsing-remitting multiple sclerosis (RRMS) and myelin oligodendrocyte glycoprotein (MOG) antibody-associated disease (MOGAD) are inflammatory demyelinating diseases of the central nervous system (CNS). Due to the shared clinical manifestations, detection of disease-specific serum antibody of the two diseases is currently considered as the gold standard for the diagnosis; however, the serum antibody levels are unpredictable during different stages of the two diseases. Herein, peripheral blood single-cell transcriptome was used to unveil distinct immune cell signatures of the two diseases, with the aim to provide predictive discrimination. Single-cell RNA sequencing (scRNA-seq) was conducted on the peripheral blood from three subjects, i.e., one patient with RRMS, one patient with MOGAD, and one patient with healthy control. The results showed that the CD19+ CXCR4+ naive B cell subsets were significantly expanded in both RRMS and MOGAD, which was verified by flow cytometry. More importantly, RRMS single-cell transcriptomic was characterized by increased naive CD8+ T cells and cytotoxic memory-like Natural Killer (NK) cells, together with decreased inflammatory monocytes, whereas MOGAD exhibited increased inflammatory monocytes and cytotoxic CD8 effector T cells, coupled with decreased plasma cells and memory B cells. Collectively, our findings indicate that the two diseases exhibit distinct immune cell signatures, which allows for highly predictive discrimination of the two diseases and paves a novel avenue for diagnosis and therapy of neuroinflammatory diseases.


Enhancing diabetic wound healing with a pH-responsive nanozyme hydrogel featuring multi-enzyme-like activities and oxygen self-supply.
Diabetic wound treating remains a challenging due to bacterial infections, oxidative stress, tissue hypoxia, and high glucose levels. Herein, a multi-enzyme-like activities nanocomposite (Mo,Fe/Cu,I-Ag@GOx) was designed and anchored to a multifunctional fluorescence hydrogel. The nanozyme gel, loaded with glucose-oxidase (GOx), exhibits intrinsic GOx, peroxidase (POD)-, oxidase (OXD)-, catalase (CAT)- and superoxide dismutase (SOD)-like activities with pH-switchable glucose-initiated cascade reaction for diabetic wound healing. In the first cascade-reaction, initiated by GOx, the nanozyme gel catalyzes glucose and O2 into gluconic acid and H2O2 to further generate superoxide anion radical (O2·-) and hydroxyl radicals (·OH) to eradicate bacteria. In the second cascade-reaction, as the wound pH changes alkalescent microenvironment, the nanozyme gel simulates SOD to transform O2·- into O2 and H2O2, and then decomposes endogenous and exogenous H2O2 into O2 via CAT-like activity to reduce oxidative stress and alleviate hypoxia. The gel by calcium ion (Ca2+) cross-linked sodium alginate (SA) and chitosan (CS) containing nanozyme was constructed with injectability, adhesion and fluorescence properties, as well as beneficial biocompatible. Importantly, the water/alcohol solubility of the nanozyme gel allows it to be used as a dressing without causing secondary injury to the wound. The multifunctional fluorescence hydrogel exhibits efficiently promote pro-angiogenesis and bacteria-infected wound healing.


Magnesium intake decreases Type 2 diabetes risk through the improvement of insulin resistance and inflammation: the Hisayama Study.
Early studies have shown that magnesium intake decreases the risk of Type 2 diabetes, but the results are still inconsistent. We prospectively examined the association between magnesium intake and incidence of Type 2 diabetes in a general Japanese population. A total of 1999 subjects without diabetes aged 40-79 years who underwent a 75-g oral glucose tolerance test were followed up prospectively for a mean of 15.6 years. During the follow-up, 417 subjects developed Type 2 diabetes. The age- and sex-adjusted incidence of Type 2 diabetes significantly decreased with increasing magnesium intake quartile levels (≤ 148.5, 148.6-171.5, 171.6-195.5 and ≥ 195.6 mg/day, P for trend = 0.01). In multivariate analyses, after adjusting for comprehensive risk factors and other dietary factors, the hazard ratio of Type 2 diabetes was 0.67 (95% CI 0.49-0.92; P = 0.01) in the third quartile and 0.63 (95% CI 0.44-0.90; P = 0.01) in the highest quartile compared with the first quartile. In addition, the risk of Type 2 diabetes was 14% lower (P = 0.04) for a 1-sd increment of log-transformed magnesium intake in the multivariate-adjusted model. In stratified analysis, there were statistically significant interactions between magnesium intake and levels of homeostasis model assessment of insulin resistance, high-sensitivity C-reactive protein or alcohol intake on the risk of Type 2 diabetes (all P < 0.05). Our findings suggest that increased magnesium intake was a significant protective factor for the incidence of Type 2 diabetes in the general Japanese population, especially among subjects with insulin resistance, low-grade inflammation and a drinking habit.


Acute Exacerbation of Interstitial Lung Disease After Cryobiopsy.
Cryotherapy has been used in treatment of lung cancer for decades. The utility of cryotechnology in diagnosis of lung diseases is emerging and gaining popularity. Cryobiopsy (CB) of the lung, when compared with conventional transbronchial forceps lung biopsy, has proposed to have a higher diagnostic yield in interstitial lung disease by providing larger biopsy specimen and less crush artifact. Acute exacerbation of interstitial lung disease (AEILD) has been well described with surgical lung biopsies and, rarely, with conventional transbronchial forceps biopsy. The incidence of AEILD after CB is not known. Here we are presenting a case of AEILD after CB.


Anti-CD11c antibody, Efalizumab attenuate ventilator-induced lung injury.
The pathophysiology of ventilator-induced lung injury (VILI) involves multiple mechanisms including inflammation and inflammatory cells infiltration. The anti-CD11c monoclonal antibody, Efalizumab has been demonstrated to inhibit the T cell activation, migration and adhesion to keratinocytes. In this study, we induced lung injury with mechanical ventilation in male Sprague-Dawley rats, the rats were divided into four groups: lung-protective ventilation (LV), injurious ventilation (HV), HV+human IgG control and HV+ Efalizumab groups. Then we detected the lung tissue wet/dry ratio, and the activity of myeloperoxidase (MPO) was determined. The concentration of protein, TNF-a, IL-6, IL-1b and MIP-2 in the BALF were detected by ELISA. The expression ICAM-1 was measured by Realtime PCR. Compared with the human IgG control treated group, the treatment of Efalizumab attenuate the ventilator-induced lung injury, including the wet/dry ratio and the activity of myeloperoxidase (MPO), meanwhile, the level of pro-inflammatory cytokines, such as TNF-a, IL-6, IL-1b and MIP-2 were decreased in the BALF of Efalizumab-treated group rats compared with the human IgG-treated control group. In addition, the histopathological index of ventilator-induced lung injury was improved after efalizumab treatment, that also reduced the recruitment of inflammatory cells into the lung, such as neutrophils. Our data suggested that Efalizumab could protect rat from ventilator-induced lung injury and improve the survival time through the inhibition of intrapulmonary inflammatory response.


Current status of global research on pericardial diseases: a bibliometric analysis of the top 100 from Web of Science.
Bibliometric studies can help guide researchers and funding bodies toward fields where more research activity is warranted. Bibliometric analyses have previously been published in many specialties and sub-specialties. Our literature search did not show a bibliometric analysis on pericardial diseases. We performed a bibliometric analysis of the top 100 cited manuscripts on pericardial diseases to identify knowledge. Bibliometric analysis is a quantitative method to assess research performance and analyze publication trends. Web of Science was searched in April 2020 to identify the top 100 cited manuscripts in pericardial diseases. Twenty-six out of the top 100 cited manuscripts were published between 2000 and 2009. These manuscripts were cited on average189 times (range: 110-743) since publication. Only two manuscripts were cited > 500 times. Among the top-ten cited manuscripts, there were 6 original articles, 1 case series, and 3 review articles. Of the 3 review articles, 2 were society guidelines. 90% of the authors had written just 1 manuscript. There were ten manuscripts with women as first authors with a significant association between gender of the first and corresponding author (odds ratio = 44, p < 0.001). Only 20% of manuscripts were funded. Most publications came from institutions in the United States (n = 40), Italy (n = 10), and Spain (n = 5). Our study provides an insight into the characteristics and quality of the highly cited literature in the field of pericardial diseases. This can be used to guide further research in the field of pericardial diseases.


Cancer-Cell Imaging Using Copper-Doped Zeolite Imidazole Framework-8 Nanocrystals Exhibiting Oxidative Catalytic Activity.
Copper-doped zeolite imidazole framework-8 (Cu/ZIF-8) was prepared and its peroxidase-like oxidative catalytic activity was examined with a demonstration of its applicability for cancer-cell imaging. Through simple solution chemistry at room temperature, Cu/ZIF-8 nanocrystals were produced that catalytically oxidized an organic substrate of o-phenylenediamine in the presence of H2 O2 . In a similar manner to peroxidase, the Cu/ZIF-8 nanocrystals oxidized the substrate through a ping-pong mechanism with an activation energy of 59.2 kJ mol-1 . The doped Cu atoms functioned as active sites in which the active Cu intermediates were expected to be generated during the catalysis, whereas the undoped ZIF-8 did not show any oxidative activity. Cu/ZIF-8 nanocrystals exhibited low cell toxicity and displayed catalytic activity through interaction with H2 O2 among various reactive oxygen species in a cancer cell. This oxidative activity in vitro allowed cancer-cell imaging by exploiting the photoluminescence emitted from the oxidized product of o-phenylenediamine, which was insignificant in the absence of the Cu/ZIF-8 nanocrystals. The results of this study suggest that the Cu/ZIF-8 nanocrystal is a promising catalyst for the analysis of the microbiological systems.


Proficient and cost-effective approaches for the prevention and treatment of venous thrombosis and thromboembolism.
Thrombosis is clearly a common cause of death in the US. It is obviously of major importance to define the aetiology of deep vein thrombosis (DVT) as (i) many of these events are preventable if appropriate therapy, dependent upon the risk factors known is utilised; (ii) appropriate antithrombotic therapy will decrease risks of recurrence; (iii) the type of defect(s) and risk(s) will determine length of time the patient should remain on therapy for secondary prevention and (iv) if the defect is hereditary appropriate family members can be assessed. Aside from mortality, significant additional morbidity occurs from DVT including, but not limited to, stasis ulcers and other sequelae of post-phlebitic syndrome. Numerous studies have provided evidence that medical patients and patients undergoing surgery or trauma are at significant risk for developing DVT, including pulmonary embolism (PE). Thus, an important task for the clinician is to prevent DVT and its complications. It is important to define risk groups where prophylaxis must be considered. The attitudes and beliefs towards prophylaxis show great regional variations. This is true for the definition of risk groups, the proportion of patients receiving prophylaxis and prophylactic modalities used. For this reason, various 'consensus conference' groups have attempted to alleviate these problems; the primary mission of consensus guidelines is to provide optimal direction to the clinician in the setting of clinical practice. If the practice guidelines generated are successful they will assist clinicians in decision-making for their patients, and they will also provide protection against unjustified malpractice actions. Therapy may be complex, as clinical studies continue to identify more effective treatments. This review includes currently accepted approaches to the treatment of DVT. The clinical course of DVT is highly dynamic. When the response to therapy is not as expected, more than one cause of DVT may be present in a patient. Treatment must address the primary coagulopathy as well as any precipitating factors. The risk of pharmacological intervention must be balanced against potential benefit. If the incidence of DVT in a given disorder is low and if the mortality rate is similarly low, therapy with an agent known to be associated with a high risk for complications, such as warfarin, would not be indicated. If DVT is seen primarily after surgery or in other high-risk situations, therapy might be limited to a fixed time period. However, if the ongoing risk of DVT remains high or if a history of recurrent DVT dictates, lifelong therapy might be indicated. The recommendations presented are based upon published controlled trials; however, indications for therapy and therapeutic agents of choice will continually evolve. By applying the principles outlined in this review, substantial cost savings, reduction in morbidity and reductions in mortality should occur.


Elevation determines the productivity of large cardamom (Amomum subulatum Roxb.) cultivars in Sikkim Himalaya.
Large cardamom (Amomum subulatum Roxb.) is an economically important cash crop that provides a livelihood option for the rural communities in Sikkim Himalaya. However, its production has declined drastically over the past few decades due to climate change and other factors affecting the livelihood of marginal cardamom-dependent farmers in the region. Climate change causes a shift in elevational distributions of mountain species, and it is pivotal to understand its effect on yield and yield-related traits for economically important plant species like large cardamom. For this, we randomly studied 41 large cardamom cultivation sites in Sikkim (India) with elevations ranging between 975 and 2069 m asl and evaluated the yield-related traits (number of capsules per spike, capsule length, capsule width, fresh capsule weight, dry capsule weight, number of seeds per locule, fresh seed weight, and dry seed weight) in five cultivars (Dzongu Golsey, Sawney, Seremna, Ramsey, and Varlangey). We observed a significant variability (P < 0.05) for morphometric traits in each of the five cultivars cultivated in different elevations. The cultivation of low-elevation cultivars like Seremna and Dzongu Golsey (suitable in elevation < 975 m) has shifted upward to mid (975-1515 m) and high-elevation (> 1515 m), while cultivation of high-elevation Ramsey cultivar (suitable in elevation > 1515 m) has shifted downward (< 1515 m). The Dzongu Golsey, Sawney, and Seremna cultivated in mid-elevation (975-1515 m) performed better in terms of yield-related traits than the same cultivars cultivated in high-elevation (> 1515 m) and showed moderate to high negative correlation between elevation and yield-related traits, indicating the negative effect of elevation on their yield. Likewise, Ramsey and Varlangey cultivated in high elevation (> 1515 m) performed better than the one cultivated in mid-elevation (975-1515 m) and depicted moderate to high positive correlation between elevation and yield-related traits, suggesting a positive influence of elevation on their yield. Although there is an elevational shift in the cultivation of large cardamom cultivars, the elevation influences the performance of the large cardamom cultivars, and it also suggests cultivating the cultivars in their suitable elevation range for better productivity.


Transcriptional activity across the Epstein-Barr virus genome in Raji cells during latency and after induction of an abortive lytic cycle.
We have studied the relative rate of transcription across the Epstein-Barr virus genome in the Burkitt's lymphoma cell line Raji by nuclear run-on analysis during latency and after induction of an abortive lytic cycle with 12-0-tetradecanoylphorbol 13-acetate (TPA) and 5-iodo-2'-deoxyuridine (IUdR). During latency the entire, or almost the entire, viral genome was found to be transcriptionally active to a low or intermediate extent, with some variation in activity along the genome. The fragment with the highest transcriptional activity was EcoRI J, which contains the genes encoding the small nuclear RNAs EBER1 and -2, transcribed predominantly by RNA polymerase III. An intermediate level of transcription was observed between positions 10 and 138 (kb), with areas of slightly higher activity on the large internal repeats and the left duplicated region (DL). The remaining part of the viral genome, between position 138 and the termini, and the termini and position 10 (kb) (with the exception of the EcoRI J fragment), showed very little transcriptional activity, except for the intermediately active regions carrying the righthand oriLyt (DR) and the terminal repeats. Upon induction of the viral genome with TPA and IUdR, the viral genome was transcriptionally active at a rate at least tenfold that seen during latency. Polymerases were not equally distributed along the genome after induction; the highest density was found in regions 48 to 58 kb, 82 to 84 kb, 102 to 104 kb, 118 to 122 kb and 142 to 145 kb of the viral genome. High transcriptional activity correlated with distinct transcription units in some cases, i.e. BamHI H1LF1 (DL), BamHI MLF1, BamHI ZLF1/BamHI RLF1 and BamHI X (thymidine kinase), but not in others (BamHI H2). Besides initiation of transcription, other regulatory processes such as stabilization and processing of primary transcripts may also contribute to regulation of virus gene expression. Addition of cycloheximide completely abolished the transcriptional activation of the genome mediated by TPA and IUdR.


[THE EPIDEMIOLOGICAL SITUATION OF LEISHMANIASIS IN THE RUSSIAN FEDERATION: THE FIRST VALID CASES OF LOCAL TRANSMISSION].
The authors have analyzed the data available in the literature and statistical offices and the results of their observations of the epidemiological situation of leishmaniasis in the Russian Federation (RF). In October 1991 to December 2014, the RF registered 89, leishmaniasis cases, including 83 imported cases from 24 countries and 6 local cases. Out of the 83 imported cases, there were those diagnosed with anthroponotic cutaneous leishmaniasis (n=31), zoonotic cutaneous leishmaniasis (n=20), visceral leishmaniasis (n=31), and mucocutaneous leishmaniasis (n=1). The Republic of Dagestan and Eastern Crimea are visceral Ieishmaniasis-enderpic, areas. The obtained data on the local cases of visceral leisimaniasis in Dagestan and on its sporadic cases in Crimea suggest that it is necessary to conduct epidemiological, epizootological, and entomological surveys in the above areas and to develop a system for visceral leishmaniasis epidemiological surveillance encompassing the whole package of antiepidemic measures. The effective treatment of leishmaniases requires that the RF Ministry of Health should register some current antileishmanial drugs (such as Glucantime) as soon as possible.


Role of Dorsomedial Striatum Neuronal Ensembles in Incubation of Methamphetamine Craving after Voluntary Abstinence.
We recently developed a rat model of incubation of methamphetamine craving after choice-based voluntary abstinence. Here, we studied the role of dorsolateral striatum (DLS) and dorsomedial striatum (DMS) in this incubation. We trained rats to self-administer palatable food pellets (6 d, 6 h/d) and methamphetamine (12 d, 6 h/d). We then assessed relapse to methamphetamine seeking under extinction conditions after 1 and 21 abstinence days. Between tests, the rats underwent voluntary abstinence (using a discrete choice procedure between methamphetamine and food; 20 trials/d) for 19 d. We used in situ hybridization to measure the colabeling of the activity marker Fos with Drd1 and Drd2 in DMS and DLS after the tests. Based on the in situ hybridization colabeling results, we tested the causal role of DMS D1 and D2 family receptors, and DMS neuronal ensembles in "incubated" methamphetamine seeking, using selective dopamine receptor antagonists (SCH39166 or raclopride) and the Daun02 chemogenetic inactivation procedure, respectively. Methamphetamine seeking was higher after 21 d of voluntary abstinence than after 1 d (incubation of methamphetamine craving). The incubated response was associated with increased Fos expression in DMS but not in DLS; Fos was colabeled with both Drd1 and Drd2 DMS injections of SCH39166 or raclopride selectively decreased methamphetamine seeking after 21 abstinence days. In Fos-lacZ transgenic rats, selective inactivation of relapse test-activated Fos neurons in DMS on abstinence day 18 decreased incubated methamphetamine seeking on day 21. Results demonstrate a role of DMS dopamine D1 and D2 receptors in the incubation of methamphetamine craving after voluntary abstinence and that DMS neuronal ensembles mediate this incubation. In human addicts, abstinence is often self-imposed and relapse can be triggered by exposure to drug-associated cues that induce drug craving. We recently developed a rat model of incubation of methamphetamine craving after choice-based voluntary abstinence. Here, we used classical pharmacology, in situ hybridization, immunohistochemistry, and the Daun02 inactivation procedure to demonstrate a critical role of dorsomedial striatum neuronal ensembles in this new form of incubation of drug craving.


Push-pull perfusion of the hypothalamus and the caudate nucleus in conscious, unrestrained animals.
The present results further confirm the usefulness of the push-pull cannula to study the in vivo activity of several neural systems in conscious, freely moving animals. However, it is evident that the inflammatory reaction to the local injury induced by the cannula may have different consequences in the function of a particular brain area. In the caudate nucleus, the responsiveness of the dopaminergic terminals of the nigrostriatal DA system to local infusion of AMPH was clearly reduced after the first two to three weeks post cannula implant. In contrast, in the hypothalamus of the rat and the rabbit, the spontaneous fluctuating activity of the LHRH pulse generator remained unaffected during a two to three month period when several perfusions could be performed in the same animal. Further studies will be required to establish the reasons for such marked differences in response to injury of these two brain structures. Various neurochemicals can be measured simultaneously from the hypothalamus of conscious rabbits. The present results indicate that NE hypothalamic impulse flow increases during the afternoon whereas the impulse flow of DA (as estimated by DOPAC output) decreases or does not change. In rabbit No. 2, simultaneous measurement of hypothalamic NE release and LHRH release indicates an absence of correlation. In sharp contrast, the simultaneous measurement of prostaglandin E2 release and LHRH release from the hypothalamus of rabbit 2-ER revealed a closer association between these two neuronal events. Apparently the push-pull technique can be used to examine long-term seasonal as well as circadian (extended 24 hr perfusion) spontaneous changes in the activity of the LHRH pulse generator in individual animals. It seems that seasonal changes characterize the function of the rabbit LHRH pulse generator. Lastly, physiological doses of progesterone infused for short times directly into the hypothalamus of female rabbit stimulate the LHRH pulse generator of this species with marked increases in the amplitude of the LHRH signal. In conclusion, the PPP technique coupled to HPLC-EC and RIA procedures to measure localized release of neurotransmitters, metabolites, and neuropeptides simultaneously in the same perfusate samples from discrete brain areas appears to be a powerful technique in furthering our knowledge of the in vivo local neurochemistry of the brain of conscious, freely moving animals.


Strategies for High Grafting Efficiency of Functional Ligands to Lipid Nanoemulsions for RGD-Mediated Targeting of Tumor Cells In Vitro.
Lipid nanoemulsions (LNEs) are promising nanocarriers for delivering high payloads of lipophilic molecules. Nonetheless, the dynamic nature at their aqueous interfaces results in poor surface chemistry and thus ligand functionalization can be challenging. Herein, two independent strategies, postconjugation and preconjugation, were explored to prepare LNEs grafted covalently with model ligands, fluorescein dye and RGD peptide, respectively. Fluorescein was successfully conjugated with high grafting efficiency to an amine-functionalized lipid nanoemulsion (NH2-LNE) as determined by spectrophotometric analysis. First, we formulated NH2-LNEs by a low-energy spontaneous emulsification technique in the presence of oleylamine (OA) within the oily core of the nanodroplets, thus creating primary amine-reactive sites at the oil/water interface. These amines were used to incorporate fluorescein, yielding fluorescent LNEs with grafting efficiencies of 33, 69, and 69% at NH2-LNEs with [OA]oil = 0.18, 0.34, and 0.49 M, respectively. We also developed RGD-labeled LNEs (RGD-LNEs) and evaluated the nanomaterial with model cell lines that overexpress αVβ3 integrins on their surfaces. To this end, we initially synthesized an RGD-Oleate fatty acid-peptide conjugate by solid-phase synthesis. The lipophilic segment of this conjugate readily embedded into the oily core of the LNE, and the hydrophilic head (RGD moiety) was oriented toward the LNE interface. In vitro cytotoxicity and cellular uptake studies were undertaken on different cancer cell lines including HaCaT human umbilical vein endothelial cells (HUVECs), MCF-7, and U-87 MG and compared to uptake experiments with RAW 264.7 macrophages. Confocal imaging and flow cytometry showed that RGD-LNEs were preferentially taken up by all of the tumor cell lines but showed very slight accumulation in RAW macrophages. Unmodified LNE controls did not show any appreciable cellular uptake. This work provides a simple and reliable methodology for the incorporation of multiple ligands on a single surface to facilitate active tumor targeting with LNE-based drug/imaging carriers for theranostic applications.


Berezinskii-Kosterlitz-Thouless transition from neural network flows.
We adopt the neural network (NN) flow method to study the Berezinskii-Kosterlitz-Thouless (BKT) phase transitions of the two-dimensional q-state clock model with q≥4. The NN flow consists of a sequence of the same units that proceed with the flow. This unit is a variational autoencoder trained by the data of Monte Carlo configurations in unsupervised learning. To gauge the difference among the ensembles of Monte Carlo configurations at different temperatures and the uniqueness of the ensemble of NN-flow states, we adopt the Jensen-Shannon divergence (JSD) as the information-distance measure "thermometer." This JSD thermometer compares the probability distribution functions of the mean spin value of two ensembles of states. Our results show that the NN flow will flow an arbitrary spin state to some state in a fixed-point ensemble of states. The corresponding JSD of the fixed-point ensemble takes a unique profile with peculiar features, which can help to identify the critical temperatures of BKT phase transitions of the underlying Monte Carlo configurations.


Species-driven phases and increasing structure in early-successional plant communities.
Successional phases describe changes in ecological communities that proceed in steps rather than continuously. Despite their importance for the understanding of ecosystem development, there still exists no reliable definition of phases and no quantitative measure of phase transitions. In order to obtain these data, we investigated primary succession in an artificial catchment (6 ha) in eastern Germany over a period of 6 years. The data set consists of records of plant species and their cover values, and initial substrate properties, both from plots in a regular grid (20 m × 20 m) suitable for spatial data analysis. Community assembly was studied by analyses of species co-occurrence and nestedness. Additionally, we correlated lognormal and log series distributions of species abundance to each community. We here introduce a new general method for detection of successional phases based on the degree of transient spatial homogeneity in the study system. Spatially coherent vegetation patterns revealed nonoverlapping partitions within this sequence of primary succession and were characterized as two distinct ecological phases. Patterns of species co-occurrence were increasingly less random, and hence the importance of demographic stochasticity and neutral community assembly decreased during the study period. Our findings highlight the spatial dimension of successional phases and quantify the degree of change between these steps. They are an element for advancing a more reliable terminology of ecological successions.


Diagnostic Accuracy of Ga-68-HBED-CC-PSMA-Ligand-PET/CT before Salvage Lymph Node Dissection for Recurrent Prostate Cancer.
Background: By targeting the prostate-specific membrane antigen (PSMA) on prostate cancer (PCa) cells PSMA-PET/CT shows great potential in locating the site of biochemical recurrence even at low PSA (Prostate-specific antigen)-levels. Accurate imaging of PCa recurrent lymph node metastases (LNM) is crucial for metastases directed therapies such as salvage-lymph node dissection (salvage-LND). Objective: To evaluate the diagnostic accuracy of PSMA-PET/CT for detection of affected lymph-node regions at salvage-LND for nodal recurrence of PCa. Design, setting and participants: 30 patients with the suspicion of exclusively nodal PCa-relapse after primary therapy underwent a template pelvic and/or retroperitoneal salvage-LND after whole body 68-Ga-PSMA-PET/CT. The diagnostic accuracy of PET/CT was evaluated in comparison to the histopathology of 965 resected lymph nodes (LN) dissected from 68 main regions (pelvic left/right, retroperitoneal) and 289 subregions (common iliac, external iliac, obturator, internal iliac, presacral, aortic-bifurcation, aortal, caval). LNM and tumor deposits in LNM were measured bidimensionally in the histopathology. PSMA-expression was analyzed by immunohistochemistry in LNM. Results: LNM were present in 11.4% of the resected LN (110/965) resulting in 45 positive main regions and 85 positive subregions. PET/CT was true positive in 41 main regions and 69 subregions. Three PET-negative main regions and 16 PET-negative subregions finally contained LNM, the majority of these false negative subregions (13/16) were in neighboring regions of true-positive subregions. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy were: main region-based 93.2%, 100%, 100%, 88.9% and 95.6%, subregion-based 81.2%, 99.5%, 98.6%, 92.7 and 94.1%. Median short diameters of tumor deposits in LNM resected from false-negative subregions (1.3 mm) were significantly smaller than in LNM removed from true-positive subregions (5.5 mm, p<0.0001). Based on anatomical subregions containing just one LNM, the necessary short diameter of tumor deposits in LNM required to reach a detection rate of 50% and 90% was estimated to be ≥ 2.3 mm and ≥ 4.5 mm, respectively. Conclusion: In men with biochemical PCa-relapse and positive PSMA-PET/CT, PET/CT detects metastatic affected anatomical regions with high accuracy at a main region and at a subregion-level. If the decision for salvage-LND is prompted by a positive PSMA-PET/CT, the size of metastases is crucial for accurate detection of affected regions. All LNM showed a clear PSMA-expression in the immunohistochemistry. Further studies need to investigate how to translate the high anatomical correlation observed between PET/CT and surgical findings into optimal approaches for target salvage-LND.


Automatic elasticity measurement of single cells using a microfluidic system with real-time image processing.
The mechanical properties of cells are closely related to their physiological states and functions. Due to the limitations of conventional cell elasticity measurement technologies such as low throughput, cell-invasiveness, and high cost, microfluidic systems are emerging as powerful tools for high-throughput cell mechanical property studies. This paper introduces a microfluidic system to automatically measure the elastic modulus of single cells in real time. The system integrated a microfluidic chip with a microchannel for cell constriction, a pressure pump, a precision differential pressure sensor, and a program for online analysis of cell deformation. The program used a fast U-net to segment cell images and measure protrusion length during cell deformation. Subsequently, the cell elasticity was determined in real-time based on the deformation and required pressure using the power law rheological model. Finally, Young's modulus of BMSCs, Huh-7 cells, EMSCs, and K562 cells was measured as 25.13 ± 15.19 Pa, 69.74 ± 92.01 Pa, 54.50 ± 59.31 Pa and 58.43 ± 27.27 Pa, respectively. The microfluidic system has significant application potential in the automated evaluation of cell mechanical properties.Clinical Relevance-The technique in this paper may be used for the automatic and high throughput study of the stiffness of cells, such as stem cells and cancer cells. The stiffness data may contribute to stem cell therapy and cancer research.