Natural history of experimental histoplasmic choroiditis in the primate. II. Histopathologic features.
The histopathologic features of experimental multifocal histoplasmic choroiditis in the primate are presented. Eyes were studied at various time periods from 30 days to 3 years after intracarotid injection with live Histoplasma capsulatum organisms. The acute lesions produced resolved into four distinct resolution patterns (chorioretinal scars, retinal pigment epithelial defects, subclinical lesions, and "disappearing" lesions) whose histopathologic features were studied. Organisms could not be demonstrated by culture or special stains in lesions present more than 6 weeks. A feature common to all eyes studied, including those whose lesions were clinically inactive or had clinically "disappeared," was the persistence of small foci of lymphocytes in the choroid. Damage to Bruch's membrane was observed in some specimens.


[Clinical study of the month. Attempt at preservation of B cells during the initial phase of type 1 diabetes: negative results with ultralente insulin, but promising results with an anti-CD3 monoclonal antibody].
Type 1 diabetes is an autoimmune disease leading to a progressive exhaustion of the insulin secretion and a destruction of the B-cells. Attempts of preservation of insulin-producing B-cells can be performed at an early, most often silent, stage of the disease in well-selected at high risk subjects or during the period immediately following the clinical diagnosis based upon classical signs of hyperglycaemia. In the "Diabetes Prevention Trial-Type 1", the prophylactic subcutaneous administration of low-dose ultralente insulin was not able to prevent the development of type 1 diabetes nor to preserve residual insulin secretion in young relatives at very high-risk of diabetes, selected upon genetic, immunological and metabolic criteria. In contrast, a pilot randomized trial shows that a treatment with a nonactivating humanized monoclonal antibody against CD3 mitigates the deterioration in insulin production and improves metabolic control, with lower dose of exogenous insulin, during the first year of type 1 diabetes mellitus in 9 out of 12 treated patients. Besides a better understanding of the natural history of the disease, these clinical trials open new perspectives for prevention of type 1 diabetes mellitus, currently assessed by the Belgian Diabetes Registry.


Immunogenicity risk assessment for biotherapeutics through in vitro detection of CD134 and CD137 on T helper cells.
Biotherapeutics, which are biologic medications that are natural or bioengineered products of living cells, have revolutionized the treatment of many diseases. However, unwanted immune responses still present a major challenge to their widespread adoption. Many patients treated with biotherapeutics develop antigen-specific anti-drug antibodies (ADAs) that may reduce the efficacy of the therapy or cross-react with the endogenous counterpart of a protein therapeutic, or both. Here, we describe an in vitro method for assessing the immunogenic risk of a biotherapeutic. We found a correlation between clinical immunogenicity and the frequency with which a biotherapeutic stimulated an increase in CD134, CD137, or both cell surface markers on CD4+ T cells. Using high-throughput flow cytometry, we examined the effects of 14 biotherapeutics with diverse rates of clinical immunogenicity on peripheral blood mononuclear cells from 120 donors with diverse human leukocyte antigen class II-encoding alleles. Biotherapeutics with high rates of ADA development in the clinic had higher proportions of CD4+ T cells positive for CD134 or CD137 than biotherapeutics with low clinical immunogenicity. This method provides a rapid and simple preclinical test of the immunogenic potential of a new candidate biotherapeutic or biosimilar. Implementation of this approach during biotherapeutic research and development enables rapid elimination of candidates that are likely to cause ADA-related adverse events and detrimental consequences.


The future of medical education: a Canadian environmental scan.
One hundred years after the Flexner report remade medical education in North America, many countries are reviewing the purpose and organisation of medical education. In Canada, a national study is being undertaken to define important issues and challenges for the future of medical education. The objectives of this paper are to describe the process of conducting an empirical environmental scan at a national level, and to present the research findings of this scan. Thirty national key informant interviews were conducted, transcribed and coded to identify key themes. Interview data were triangulated with data sourced from 34 commissioned literature reviews and a series of national focus groups. Ten key issues or priorities were identified and used to generate detailed review papers used by the Association of Faculties of Medicine of Canada to create a blueprint for the evolution of medical education. The new priorities have major implications for areas ranging from admissions, curriculum content, educational process and the need to articulate the purpose and responsibilities of medical schools in society. This research provides a case study of how an empirical research approach can be used to identify and validate priorities for changes in medical education at a national level. This approach may be of interest in other countries.


Results of Casting in Severe Curves in Infantile Scoliosis.
Previous work has demonstrated best results for casting in infantile scoliosis when the curves are small and the child begins casting under 2 years of age. This study examines if casting can delay the need for growth friendly instrumentation in severe curves (50 to 106 degrees) and how the comorbidities of syrinx or genetic syndromes affected outcomes. All children undergoing casting for scoliosis at a single institution over an 8-year period were examined. Inclusion criteria included initial curve at first casting of ≥50 degrees, age ≤3 years at the start of casting, and a minimum follow-up of 3 years. Of 148 children undergoing casting during this period, 44 met our inclusion criteria. All children underwent magnetic resonance imaging. Ten children with a syrinx were identified. Ten children had known genetic syndromes (2 who also had a syrinx). The 26 children without these comorbidities were considered idiopathic. Curve magnitude ranged from 50 to 106 degrees. Nine of the 26 (35%) children in the children with idiopathic curves demonstrated resolution of their curves, while only 3 of the remaining 18 (17%) did. Of the children that did not have resolution of their curves, 14 were maintained over the entire follow-up period to within 15 degrees of their initial curve and 13 were improved 15 degrees or more. Only 5 children had an increase of 15 degrees or more over the follow-up period and 4 of these have undergone growth friendly instrumentation after a mean delay from initial cast of 71 months (range, 18 to 100 mo). This study demonstrates that even in severe curves, casting was effective in delaying instrumentation in all cases, and led to curve resolution of the curves in 12 of 44 children. Level III-case control study.


A linear model of acoustic-to-facial mapping: model parameters, data set size, and generalization across speakers.
The relationship between acoustic and visual speech is important for understanding speech perception, but it also forms the basis behind a type of facial animator, which can predict facial motion during speech given an acoustic input. This relationship was examined by revisiting a linear transformation model of audio-visual speech production. A mathematical model is constructed whereby the visual aspect of speech is reproduced from the acoustic signal via a linear transformation. Unlike previous studies in this area, this paper will address specific aspects of the model as related to the effects of window size for acoustic framing and the critical size of the training set. On average, facial motion is predicted with a correlation of 0.70 to the recorded motion, when the model is trained and then tested on the same subject. This is comparable to previous studies using either similar or different model approaches. Using a model trained on other subjects and then applying it to a new subject resulted in a prediction correlation of 0.65. Furthermore, acoustic windows of 100 ms and a data set of approximately 40 sentences are required for maximum predictability. The results are interpreted in terms of the underlying assumptions of the model.


Glutamine synthetase gene knockout-human embryonic kidney 293E cells for stable production of monoclonal antibodies.
Previously, it was inferred that a high glutamine synthetase (GS) activity in human embryonic kidney (HEK) 293E cells results in elevated resistance to methionine sulfoximine (MSX) and consequently hampers GS-mediated gene amplification and selection by MSX. To overcome this MSX resistance in HEK293E cells, a GS-knockout HEK293E cell line was generated using the CRISPR/Cas9 system to target the endogenous human GS gene. The GS-knockout in the HEK293E cell line (RK8) was confirmed by Western blot analysis of GS and by observation of glutamine-dependent growth. Unlike the wild type HEK293E cells, the RK8 cells were successfully used as host cells to generate a recombinant HEK293E cell line (rHEK293E) producing a monoclonal antibody (mAb). When the RK8 cells were transfected with the GS expression vector containing the mAb gene, rHEK293E cells producing the mAb could be selected in the absence as well as in the presence of MSX. The gene copies and mRNA expression levels of the mAb in rHEK293E cells were also quantified using qRT-PCR. Taken together, the GS-knockout HEK293E cell line can be used as host cells to generate stable rHEK293E cells producing a mAb through GS-mediated gene selection in the absence as well as in the presence of MSX.


Identification of a Novel Gibberellin (GA85) in Very Young Seedlings of Brassica campestris cv. Tobin.
A new gibberellin (GA) was identified from extracts of cotyledons of 7 day-old canola seedlings (Brassica campestris cv. Tobin). This GA is 12α-hydroxy-GA1 and has been assigned the trivial name of GA85. Isolation was monitored by the Tan-ginbozu dwarf rice micro-drop assay after each high-performance liquid chromatography (HPLC) step. Identification was based on Kovats retention index (KRI) and the mass spectrum of the methyl ester, trimethylsilyl ether (MeTMSi) derivative after analysis by gas chromatography-mass spectrometry (GC-MS) in comparison with an authentic sample of 12α-hydroxy-GA1. Based on quantitation by the dwarf rice micro-drop assay, GA85 is one of the major biologically active GAs in cotyledons of young canola seedlings.


Third-party social evaluation of humans by monkeys.
Humans routinely socially evaluate others not only following direct interactions with them but also based on others' interactions with third parties. In other species, 'eavesdropping' on third-party interactions is often used to gain information about foraging or mating opportunities, or others individuals' aggressiveness or fighting ability. However, image scoring for potential cooperativeness is less well studied. Here we ask whether a non-human primate species, tufted capuchin monkeys (Cebus apella), socially evaluates humans after witnessing third-party interactions involving a helpful intervention or failure to help. We find that the monkeys accept food less frequently from those who persistently reject another's requests for help. This negative social evaluation effect is robust across conditions, and tightly linked to explicit refusal to help. Evaluation of potential helpfulness based on third-party interactions may thus not be unique to humans.


Exploring employment preferences of nurses to improve retention in rural China.
The scarcity of nurses in rural China deserves serious consideration in light of the widening gap in health outcomes between rural and urban populations. The outflow of primary care nurses suggests the value of examining their employment preferences to identify possible effective incentives for their retention. Our research used discrete choice experiment methodology designed to quantitatively measure the relative importance of various job attributes valued by primary care nurses using conditional logit and mixed logit models. Willingness to pay and uptake rates were also calculated. A total of 1744 registered nurses in township health centres were surveyed. Participants considered all the job attributes included in the study to be significant, with pay increases demonstrating the greatest potential to improve the retention of nurses. Among the non-monetary aspects of work, nurse-patient relationships, bianzhi (positions allocated to each unit by the Chinese government with special benefits) and educational opportunities ranked highest in importance for respondents. Responses to hypothetical policy proposals suggest that incentive packages are also likely to be effective. Policy making regarding rural nurses' retention is an important undertaking that requires an evidence-based approach. Our findings provide a range of policy options that shed light on effective retention strategies. Tailoring policy incentives to primary care nurses is also necessary.


Fatal post-transplant lymphoproliferative disorder following allogeneic bone marrow transplantation for aplastic anemia.
Post-transplant lymphoproliferative disorder (PTLD) constitutes a serious complication of allogeneic bone marrow transplantation. We describe a case of PTLD in a twenty-six year-old male treated with bone marrow transplantation for aplastic anemia of unknown cause. The patient received unmanipulated marrow graft from his HLA-matched brother. Fifty-one days post transplant he developed progressive enlargement of cervical lymph nodes, followed by hepatosplenomegaly and generalized lymphadenopathy. Polymorphic PTLD was diagnosed basing on the lymph node histopathology, positive EBV detection, flow cytometry and IgH rearrangement studies proving monoclonality (capillary electrophoresis with ABI PRISM 310 Genetic Analyzer). There was no response to anti-CD20 antibody, cessation of immunosuppression, donor lymphocyte infusion and cytostatic therapy. The patient died on the 65th day of multiple organ failure. We discuss the diagnostics and management of PTLD in the setting of bone marrow transplantation.


Localization and Ordering of Lipids Around Aquaporin-0: Protein and Lipid Mobility Effects.
Hydrophobic matching, lipid sorting, and protein oligomerization are key principles by which lipids and proteins organize in biological membranes. The Aquaporin-0 channel (AQP0), solved by electron crystallography (EC) at cryogenic temperatures, is one of the few protein-lipid complexes of which the structure is available in atomic detail. EC and room-temperature molecular dynamics (MD) of dimyristoylglycerophosphocholine (DMPC) annular lipids around AQP0 show similarities, however, crystal-packing and temperature might affect the protein surface or the lipids distribution. To understand the role of temperature, lipid phase, and protein mobility in the localization and ordering of AQP0-lipids, we used MD simulations of an AQP0-DMPC bilayer system. Simulations were performed at physiological and at DMPC gel-phase temperatures. To decouple the protein and lipid mobility effects, we induced gel-phase in the lipids or restrained the protein. We monitored the lipid ordering effects around the protein. Reducing the system temperature or inducing lipid gel-phase had a marginal effect on the annular lipid localization. However, restraining the protein mobility increased the annular lipid localization around the whole AQP0 surface, resembling EC. The distribution of the inter-phosphate and hydrophobic thicknesses showed that stretching of the DMPC annular layer around AQP0 surface is the mechanism that compensates the hydrophobic mismatch in this system. The distribution of the local area-per-lipid and the acyl-chain order parameters showed particular fluid- and gel-like areas that involved several lipid layers. These areas were in contact with the surfaces of higher and lower protein mobility, respectively. We conclude that the AQP0 surfaces induce specific fluid- and gel-phase prone areas. The presence of these areas might guide the AQP0 lipid sorting interactions with other membrane components, and is compatible with the squared array oligomerization of AQP0 tetramers separated by a layer of annular lipids.


Healing High-Loading Sulfur Electrodes with Unprecedented Long Cycling Life: Spatial Heterogeneity Control.
Self-healing capability helps biological systems to maintain their survivability and extend their lifespan. Similarly, self-healing is also beneficial to next-generation secondary batteries because high-capacity electrode materials, especially the cathodes such as oxygen or sulfur, suffer from shortened cycle lives resulting from irreversible and unstable phase transfer. Herein, by mimicking a biological self-healing process, fibrinolysis, we introduced an extrinsic healing agent, polysulfide, to enable the stable operation of sulfur microparticle (SMiP) cathodes. An optimized capacity (∼3.7 mAh cm-2) with almost no decay after 2000 cycles at a high sulfur loading of 5.6 mg(S) cm-2 was attained. The inert SMiP is activated by the solubilization effect of polysulfides whereas the unstable phase transfer is mediated by mitigated spatial heterogeneity of polysulfides, which induces uniform nucleation and growth of solid compounds. The comprehensive understanding of the healing process, as well as of the spatial heterogeneity, could further guide the design of novel healing agents (e.g., lithium iodine) toward high-performance rechargeable batteries.


Degenerative meniscus: Pathogenesis, diagnosis, and treatment options.
The symptomatic degenerative meniscus continues to be a source of discomfort for a significant number of patients. With vascular penetration of less than one-third of the adult meniscus, healing potential in the setting of chronic degeneration remains low. Continued hoop and shear stresses upon the degenerative meniscus results in gross failure, often in the form of complex tears in the posterior horn and midbody. Patient history and physical examination are critical to determine the true source of pain, particularly with the significant incidence of simultaneous articular pathology. Joint line tenderness, a positive McMurray test, and mechanical catching or locking can be highly suggestive of a meniscal source of knee pain and dysfunction. Radiographs and magnetic resonance imaging are frequently utilized to examine for osteoarthritis and to verify the presence of meniscal tears, in addition to ruling out other sources of pain. Non-operative therapy focused on non-steroidal anti-inflammatory drugs and physical therapy may be able to provide pain relief as well as improve mechanical function of the knee joint. For patients refractory to conservative therapy, arthroscopic partial meniscectomy can provide short-term gains regarding pain relief, especially when combined with an effective, regular physiotherapy program. Patients with clear mechanical symptoms and meniscal pathology may benefit from arthroscopic partial meniscectomy, but surgery is not a guaranteed success, especially with concomitant articular pathology. Ultimately, the long-term outcomes of either treatment arm provide similar results for most patients. Further study is needed regarding the short and long-term outcomes regarding conservative and surgical therapy, with a particular focus on the economic impact of treatment as well.


Caries experience in 15-year-old school children in Bangalore with inherited taste sensitivity levels to 6-n-propylthiouracil: an observational study.
The relationship between sugar consumption and caries has been researched extensively for many years. The individual drive for the consumption of sweet food is controlled by a variety of biologic, psychologic, and sociologic factors. Sweet preference has been linked to bitter taste sensitivity to 6-n-propylthiouracil, a heritable trait. The present study investigates the association between inherited taste sensitivity to 6-n-popylthiouracil and dental caries experience in 15-year-old school children. Two hundred 15-year-old school children from randomly selected schools in Bangalore, India, participated in the study. Data were collected regarding demographic factors and socioeconomic status by personal interviews with the children. Sensitivity to 6-n-propylthiouracil was determined using Tepper filter paper method, and the responses were recorded on the labeled magnitude scale. Each child was examined thoroughly for oral hygiene status and caries experience. Appropriate statistical tests, such as chi-squared test, Fisher's exact test, and Kruskal-Wallis test, were used to analyze the data. Non-tasters were significantly associated with caries experience (P < 0.001). The decayed component in non-tasters (3.43 ± 2.18) was statistically significant when compared to tasters (2.86 ± 1.58 medium tasters, 1.50 ± 0.55 supertasters). After all associated factors were controlled, 6-n-propylthiouracil taste status was the only independent variable significantly related to overall caries experience.


Multiple fibromyxomas of the jaws: A case report.
Fibromyxoma of the jaw is a rare benign mesenchymal odontogenic tumor with locally aggressive behavior. In the present report, a 13-year-old female patient presented to our university hospital with delayed eruption of some of her teeth. A panoramic radiograph taken at the initial examination revealed four pericoronal radiolucencies related to the four third molars. Thereafter, a magnetic resonance imaging (MRI) examination was performed. After the surgical removal of these molars, the microscopic examination diagnosed the four lesions as fibromyxomas. Here, we have discussed the clinical, panoramic radiography, MRI, and histopathological findings of the case.


A survey of US dental practices' use of social media.
Social media is becoming the way for businesses, including health care professionals, to communicate with consumers. The study examines the use of social media by dental practices in the US. An electronic survey was sent to 22,682 dentists in the United States. The survey consisted of questions related to the use of social media in the dental practice. Data was analyzed using descriptive statistics and comparisons were made using a Fisher's exact test. A total of 573 responses were received. Social media was used in 52% of dental practices, the most common being Facebook. The use of social media was most commonly for marketing purposes (91%). Dentists younger than 45 years old were more likely to use social media in their practice than dentists 45 years or older (p > 0.001). Dental practices actively use social media for marketing and communication. Many dentists are unsure how to measure the success of social media in their practice. Additional research is needed to measure the success of social media in a dental practice. Social media is a common way practices market and interact with their patients. There are some difficulties in determining what appropriate content for social media is and how to evaluate the success.


Extrahepatic bile duct malignancies.
Biliary tract malignancies belong to very aggressive malignancies of the gastrointestinal tract. The only radical treatment is surgical resection which is possible only in a limited number of cases due to late diagnosis. The aim of this report was to present the experience of our own department with the diagnosis and treatment of these tumours. In the years 2005-2021 radical (R0) resection was performed in 27 (28.4%) patients, the same number were managed only symptomatically and in 41 (43.2%) patients we used biliary stenting and external-internal drainage as the definitive procedure. Adjuvant oncological treatment was indicated in 16 (59.3%) of the radically operated and 49 (72.1%) of the non-operated patients. Median overall survival and median progression-free survival in the operated patients were 19.9 months and 15.7 months, respectively. Overall survival in the operated patients was significantly better (p.


luxAB gene fusions with the arsenic and cadmium resistance operons of Staphylococcus aureus plasmid pI258.
pC101, a novel shuttle vector between Escherichia coli and Staphylococcus aureus carrying the lux genes encoding luciferase from Vibrio harveyi, selectable ampicillin and chloramphenicol markers and origins of replication for Gram-negative and Gram-positive bacteria has been constructed. The inducibility of the arsenic and cadmium operon from S. aureus plasmid pI258 to different ions has been tested in E. coli and in S. aureus with two fusions in pC101: an arsB-luxAB and a cadA-luxAB transcriptional gene fusion. Patterns of induction are influenced by the host strain and are slightly different from previous reports using the blaZ gene as reporter gene.


The species of Psomizopelma Gibson (Hymenoptera: Eupelmidae).
The species of the New World genus Psomizopelma Gibson (Hymenoptera: Chalcidoidea: Eupelmidae: Eupelminae) are revised. Five species are recognized, including one species with macropterous females, P. macropterum n. sp. (♀: Belize, Bolivia, Brazil, Colombia, Costa Rica, Ecuador, French Guiana, Panama, Trinidad), and four species with brachypterous females, P. albiclava n. sp. (♀ ♂: Brazil), P. brachypterum Gibson, 1995 (♀ ♂: USA, Florida), P. fuscum n. sp. (♀: Dominican Republic), and P. metallicum n. sp. (♀: USA, Florida). The species are illustrated through macrophotography and keyed.


A systematic review of arterial stiffness, wave reflection and air pollution.
Arterial stiffening is associated with increased cardiovascular risk. Whether exposure to relatively high levels of air pollution is associated with arterial stiffening is unclear. We aimed to assess the association between exposure to major air pollutants and arterial stiffening. PubMed, SCOPUS and Web of Science databases (through 31 January 2017) were searched using a combination of terms related to exposure to gaseous [nitrogen dioxide (NO2), nitrogen oxides (NOx) and sulphur dioxide (SO2)] or particulate matter pollutants (PM2.5, PM10 and PM10-2.5), arterial stiffness (pulse wave velocity) and reflected waves (augmentation index, augmentation pressure). Pertinent information were extracted from selected studies. In this systematic review were included 8 studies with available data on air pollution and arterial stiffness/reflected waves parameters (8 studies explored the effects of exposure to particulate matter pollutants, 3 studies the effects of exposure to gaseous pollutants); seven of them reported increased arterial stiffness/reflected waves after exposure to air pollution (6 of 8 studies after particulate matter pollutants; 2 of 3 studies after gaseous pollutants). Arterial stiffness and reflected waves were increased in the majority of the studies after both short- and long-term exposure to air pollutants. In conclusion, available evidence supports an association of main air pollutants with increased arterial stiffness and reflected waves. This finding may have implications for population-based strategies for the reduction of arterial stiffness, a vascular biomarker and an intermediate endpoint for cardiovascular disease.


[Simulator for Surgical Training].
Dr. William Halsted started residency-based surgical training at Johns Hopkins University in 1889. On the job training(OnJT) conducted there is an in-hospital training that residents imitate what senior doctors are doing with their patients. [ See one, do one, teach one] is the word briefly expressing this traditional training. Since then all surgeons learned surgery from the mentors and senior doctors in this way. In this OnJT, surgeons spent many years and years and accumulated experience and tried to crime up step by step from beginners to experts. Off-the-job training (OffJT) is, in other words, "planned experience". In OffJT, surgeons can experience every kind of skill by simulation. They can experience rare but critical situations, repeat the procedures while experiencing failures many times in trial and error fashion without putting any risk to the patient. In this report, currently available simulators and when and how to utilize them are introduced. Dry lab or synthetic model, wet lab or animal tissue/organ, live animal, cadaver and virtual simulation are presented and discussed.


Genetic variations in ABCB1 and CYP3A5 as well as sex influence quinine disposition among Ugandans.
Quinine is one of the most effective antimalarial drugs, although its clinical use is limited as a result of its narrow safety margin. Quinine is a substrate of the polymorphic p-glycoprotein and CYP3A4/3A5. This study aimed to examine the effects of genetic variations in ABCB1 and CYP3A5 genes, sex, demographic, and biochemical variables (serum albumin, creatinine, alanine aminotransferase and albumin) on quinine disposition among Ugandans. Quinine (600 mg) was orally administered to 140 healthy volunteers. Quinine and its metabolite 3-hydroxyquinine concentrations were determined from 16-hour postdose plasma by high-performance liquid chromatography. CYP3A5 activity was measured using quinine/3-hydroxyquinine ratio (metabolic ratio). Genotyping for a total of 20 single nucleotide polymorphisms in ABCB1 (n = 13) and CYP3A5 (n = 7) was done using Taqman and minisequencing on microarray. There were 20.5- and 13-fold variations in body weight-adjusted plasma quinine concentrations (mean +/- standard deviation, 5.26 +/- 2.5 mumol/L; range, 0.88-18.10 mumol/L) and quinine-to-3-hydroxyquinine metabolic ratio (mean +/- standard deviation, 7.68 +/- 3.3 mumol/L; range, 1.66-22.3 mumol/L), respectively. Weight-adjusted plasma quinine concentration was significantly influenced by sex and ABCB1 haplotype. There was a significant sex difference in quinine metabolic ratio, women being faster metabolizers than men (P = 0.01). CYP3A5 genotype/haplotype significantly (P = 0.03) influenced quinine disposition with a clear CYP3A5*1 gene dose effect. The result confirms that quinine disposition is influenced mainly by sex as well as by ABCB1 and CYP3A5 genotypes. Despite being fast metabolizers, women display higher quinine bioavailability than men in Uganda. This may have clinical significance in determining an individual's susceptibility to quinine-associated adverse reactions such as cinchonism.


[Mechanism of Musk and Borneol on Inflammatory of Cerebral Ischemia and Reperfusion Injury at Different Time Points of Acute Phase in Rats].
To investigate the mechanism of Musk and Borneol on cerebral ischemia and reperfusion injury at different time points of acute phase in rats. 180 rats were divided into seven groups including sham, ischemia-reperfusion after 24 h and 72 h model group, Musk 50 and 25 mg/kg groups, Borneol 50 and 25 mg/kg groups, and Xingnaojing 10 mL/kg group. Ischemia-reperfusion model was made after administration of each drug. The neurologic impairment scores at different time points after ischemia and reperfusion was evaluated, activities of cyclooxygenase (COX-2) and 5-lipoxygenase (5-LOX) in brain tissue were determined, and the expression of CysLT2 protein and mRNA in hippocampus were explored. Musk and Borneol significantly improved the neurologic impairment scores of ischemia-reperfusion injury rats, improved the pathological morphology of rats brain tissue, reduced the activities of COX-2 and 5-LOX in brain homogenates,and inhibited the expression of CysLT2 protein in hippocampus. Musk and Borneol have protective effect on inflammatory injury of acute injury in ischemia-reperfusion injury rats, the mechanism is related to inhibition the activity of COX-2 and 5-LOX in brain.


Palliative treatment and the role of surgical resection in gastric cancer.
Adenocarcinoma of the stomach is often diagnosed in the late stages of the disease. Surgical resection of all gross and microscopic disease is essential for curative treatment. Complete resection is often not achievable when patients present with advanced stage IV cancer. In the absence of symptoms, chemotherapy without resection has been the standard of care in most major centers. With improvements in response to chemotherapy and less invasive surgical approaches, patients with metastatic gastric cancer have had better survival outcomes than in the past. The challenge today when treating these patients is in defining who will benefit from more aggressive interventions. Reviewing the literature for guidance is difficult because the goals of treatment are often not clearly defined. Finding the proper balance of aggressiveness needed to extend survival while preserving and maximizing quality of life is a decision that clinicians have to make with increasing frequency. This review will attempt to provide a framework to aid in determining what role, if any, gastrectomy has in the management of patients with stage IV gastric cancer.


Lipid mediators as agonists for the resolution of acute lung inflammation and injury.
Resolution of acute lung inflammation and injury is an active process; it is not merely the absence of proinflammatory signals. Restoration of homeostasis is coordinated by specific mediators and cellular events. In response to injury and inflammatory stimuli, infiltrating leukocytes and tissue-resident cells interact to generate lipoxins (LXs), which are bioactive eicosanoids derived from arachidonic acid. In contrast to proinflammatory leukotrienes and prostaglandins, LXs display potent antiinflammatory actions. LXA(4) interacts with a G protein-coupled receptor, termed ALX, that transduces counter-regulatory signals in part via intracellular polyisoprenyl phosphate remodeling. Presqualene diphosphate (PSDP) is a polyisoprenyl phosphate in human neutrophils that is rapidly converted to presqualene monophosphate (PSMP) upon cell activation. PSDP, but not PSMP, directly inhibits phospholipase D, phosphoinositol-3 kinase, and superoxide anion generation. LXs block PSDP turnover in neutrophil membranes to prevent proinflammatory responses. Hence, LX and polyisoprenyl phosphate signaling provide a counter-regulatory circuit to promote resolution of acute lung inflammation. LXA(4) and PSDP mimetics have been prepared with potent protective actions in murine models of asthma and acute lung injury.


Polypropylene mesh implantation for hernia repair causes myeloid cell-driven persistent inflammation.
Polypropylene meshes that are commonly used for inguinal hernia repair may trigger granulomatous foreign body reactions. Here, we show that asymptomatic patients display mesh-associated inflammatory granulomas long after surgery, which are dominated by monocyte-derived macrophages expressing high levels of inflammatory activation markers. In mice, mesh implantation by the onlay technique induced rapid and strong myeloid cell accumulation, without substantial attenuation for up to 90 days. Myeloid cells segregated into distinct macrophage subsets with separate spatial distribution, activation profiles, and functional properties, showing a stable inflammatory phenotype in the tissue surrounding the biomaterial and a mixed, wound-healing phenotype in the surrounding stromal tissue. Protein mass spectrometry confirmed the inflammatory nature of the foreign body reaction, as characterized by cytokines, complement activation, and matrix-modulating factors. Moreover, immunoglobulin deposition increased over time around the implant, arguing for humoral immune responses in association with the cell-driven inflammation. Intravital multiphoton microscopy revealed a high motility and continuous recruitment of myeloid cells, which is partly dependent on the chemokine receptor CCR2. CCR2-dependent macrophages are particular drivers of fibroblast proliferation. Thus, our work functionally characterizes myeloid cell-dependent inflammation following mesh implantation, thereby providing insights into the dynamics and mechanisms of foreign body reactions to implanted biomaterials.


Multiple intra-articular injections of allogeneic bone marrow-derived stem cells potentially improve knee lesions resulting from surgically induced osteoarthritis: an animal study.
Mesenchymal stem cells (MSCs) have several properties that may support their use as an early treatment option for osteoarthritis (OA). This study investigated the role of multiple injections of allogeneic bone marrow-derived stem cells (BMSCs) to alleviate the progression of osteoarthritic changes in the various structures of the mature rabbit knee in an anterior cruciate ligament (ACL)-deficient OA model. Two months after bilateral section of the ACL of Japanese white rabbits aged nine months or more, either phosphate buffered saline (PBS) or 1 x 106 MSCs were injected into the knee joint in single or three consecutive doses. After two months, the articular cartilage and meniscus were assessed macroscopically, histologically, and immunohistochemically using collagen I and II. Within the PBS injection (control group), typical progressive degenerative changes were revealed in the various knee structures. In the single MSC injection (single group), osteoarthritic changes were attenuated, but still appeared, especially in the medial compartments involving fibrillation of the articular cartilage, osteophyte formation in the medial plateau, and longitudinal tear of the meniscus. In the multiple-injections group, the smoothness and texture of the articular cartilage and meniscus were improved. Histologically, absence or reduction in matrix staining and cellularity were noticeable in the control and single-injection groups, respectively, in contrast to the multiple-injections group, which showed good intensity of matrix staining and chondrocyte distribution in the various cartilage zones. Osteoarthritis Research Society International (OARSI) scoring showed significantly better results in the multiple-injections group than in the other groups. Immunohistochemically, collagen I existed superficially in the medial femoral condyle in the single group, while collagen II was more evident in the multiple-injections group than the single-injection group. A single injection of MSCs was not enough to restore the condition of osteoarthritic joints. This is in contrast to multiple injections of MSCs, which had the ability to replace lost cells, as well as reducing inflammation. Cite this article: Bone Joint J 2019;101-B:824-831.


Amyloid-beta (1-42) lesion of CA1 rat dorsal hippocampus reduces contextual fear memory and increases expression of microglial genes regulating neuroinflammation.
Emerging evidence indicates that the pathogenesis of Alzheimer's disease (AD) is not confined to neuronal disruptions but robustly communicates with the brain's immune system. Genome-wide analysis suggests that several genes, which increase the risk for AD, encode for factors that regulate the glial clearance of misfolded proteins and the inflammatory reaction. This study reappraises the amyloid hypothesis by focusing on the impact of neuroinflammation in a beta-amyloid model of AD, how this possibly exacerbates the disease's progression, and the correlation between genes regulating neuroinflammation (CD33 and TREM2) with post-training recall. Male Sprague-Dawley rats were used for this study, randomly divided into a vehicle group of rats (n = 40) that were infused with phosphate-buffered saline (PBS) and an Aβ(1-42) group (n = 40) that were infused with the neurotoxin Aβ(1-42) peptide. Fear conditioning test (FCT) to assess fear memory was conducted pre and post-lesion. The polymerase chain reaction was performed to determine the expression levels of CD33 and TREM2 genes. Our results show that Aβ(1-42) lesion of the rat CA1 hippocampal subregion significantly reduces contextual fear memory, and this reduction was exacerbated as the post-lesion days increased. We also observed an increase in the expression levels of CD33 and TREM2 genes in the Aβ(1-42) lesioned groups compared to their corresponding vehicle groups. Taken together, the behavioral and gene expression data provide inferential evidence that Aβ(1-42) infusion impairs contextual memory by disrupting cellular pattern separation processes in the hippocampus, thus linking neuroinflammation to specific neural circuit disruption and cognitive deficit.


Trust and distrust in low-income Michigan residents during the early COVID-19 pandemic: A qualitative study.
Trust and distrust have shaped health behaviour during the COVID-19 pandemic. Since the start of the pandemic, misinformation and polarization eroded trust across the United States. In states like Michigan, pandemic restrictions led to significant unrest. Michiganders also faced disproportionate morbidity and mortality from COVID-19 during this period. The objective of this qualitative study was to understand the individual experiences of trust in low-income Michiganders during the early COVID-19 pandemic. Twenty-four participants at or below 200% of the federal poverty line who resided in Michigan were recruited for this study. Interviews were conducted during the winter of 2020 using a formal interview guide that addressed sources of information, perceptions of risk and exposure, protective behaviours and impacts of the pandemic at home, work and in receiving healthcare. Thematic analysis showed that themes of trust and distrust emerged in multiple facets of our participants' experiences, including in the context of information sources, the behaviours of others, health, financial security, employment and overall safety. Trust and distrust in low-income communities often stemmed from significant financial and economic vulnerabilities and instability in access to healthcare that was exacerbated in the pandemic. Furthermore, participant trust was shaped by internal (e.g., relationships with others) and external (e.g., source of information, social inequity) factors that influenced their perceptions and experiences during the pandemic. Trust has played an important role in many aspects of the experiences of low-income communities during the COVID-19 pandemic. This is important for clinicians to consider as COVID-19 becomes endemic, and trust continues to impact patients' approaches to vaccines, testing and treatment options. This study was designed and conducted with the assistance and input of the members of the DECIDERS Steering Committee, a diverse statewide network of community members in Michigan. The DECIDERS team allows community members to have a voice in the design and conduct of health research, and collaborates with researchers to improve health across the state of Michigan.


From the Viewpoint of Drug Metabolism Research.
Since more than 70% of clinically used drugs are excreted from the body through metabolic processes, drug metabolism is a key determinant of pharmacokinetics, drug response and drug toxicity. Much progress has been made in understanding drug-drug interactions via the inhibition or induction of cytochrome P450s (P450, CYP), as well as the effects of genetic polymorphisms of P450s on pharmacokinetics, and this has facilitated the progress of optimized pharmacotherapy in the clinic. Now, similar information is needed for non-CYP enzymes, especially concerning Phase I enzymes, based on advanced basic and clinical studies. Recently, it was revealed that post-transcriptional regulation by microRNAs or RNA editing plays a significant role in regulating the expression of drug-metabolizing enzymes, thus conferring variability in the detoxification and metabolic activation of drugs or chemicals. Changes in the expression profile of microRNAs in tissues or body fluids can be a biomarker of drug response and toxicity; therefore, such studies could also be useful for drug repositioning. In addition, microRNAs are involved in pharmacogenetics, because single nucleotide polymorphisms in microRNA binding sites of mRNAs, or microRNAs themselves, may cause changes in gene expression. Some microRNA-related polymorphisms could be biomarkers of the clinical outcome of pharmacotherapy. In this review article, recent progress and future directions for drug metabolism studies are discussed.


The Influence of Knee Position on Ultrasound Imaging of Femoral Cartilage in Individuals with Anterior Cruciate Ligament Reconstruction.
Articular cartilage is important for knee function and can be imaged using ultrasound. The purpose was to compare femoral cartilage thickness and echo intensity (EI) measured at 90° and 140° of knee flexion and between limbs in a cohort with unilateral anterior cruciate ligament reconstruction (ACLR). We also examined associations between gait biomechanics and cartilage outcomes. Twenty-seven individuals with primary unilateral ACLR participated (12 men, 15 women; age = 22.3 ± 3.8 years; time since ACLR = 71.2 ± 47.2 months). Ultrasound was used to obtain femoral cartilage measurements. Gait outcomes included peak KFA (knee flexion angle) and peak external knee flexion moment (KFM). Cartilage outcomes were compared using a 2 (position) × 2 (limb) repeated measures ANOVA (analysis of variance). Gait and cartilage associations were assessed using linear regression. There were no position × limb interactions for any cartilage outcome (all P > 0.05). Medial (P = 0.038) and central cartilage (P < 0.001) were thicker, whereas central (P = 0.029) and lateral cartilage EI (P = 0.003) were lower when measured at 90° than those at 140° of knee flexion. Medial cartilage was thicker in the ACLR than that in the contralateral limb (P = 0.016). A larger KFM was associated with thicker medial cartilage (ΔR2 = 0.146, P = 0.021) and central cartilage (ΔR2 = 0.159, P = 0.039) measured at 140° of knee flexion in the ACLR limb but not at 90°. Findings suggest that imaging position influences cartilage thickness and EI measurements in individuals with ACLR and should be considered in study designs and clinical evaluation. A greater KFM was associated with thicker cartilage within specific portions of the distal femur.


Development of the neural crest-derived intrinsic innervation of the human lung.
The formation of neural tissue, in association with airway smooth muscle (ASM), is a feature of normal lung development and function. Intrinsic neuronal tissue has recently been shown, in animal models, to be derived from neural crest cells (NCC). Since defects in NCC development underlie a range of disease states (neurocristopathies), it is important to determine the spatiotemporal development of NCC in the human lung, as defects in their development could have pathophysiologic implications. The aims of this study were to: (1) establish a time course for the formation of ASM and neural tissue within the embryonic and fetal human lung, (2) investigate whether intrinsic neural tissue within the lung is derived from NCC, and (3) gain insight into the possible signaling mechanisms underlying the development of the intrinsic lung innervation. Using human lung tissue from Weeks 6 to 12 of gestation, we analyzed the formation of ASM, NCC, neuronal and glial tissue, and the expression of Gfralpha1, a receptor component of the RET (rearranged during transfection) tyrosine kinase signaling pathway. Our results showed that NCC accumulated along the branching airways, in close association with the ASM, and differentiated into neurons and glia. Neural crest-derived neural tissue within the lung strongly expressed membrane-bound Gfralpha1, and soluble Gfralpha1 was expressed within the lung mesenchyme, but only at early developmental stages. Together these findings indicate that the intrinsic innervation of the human lung is derived from the neural crest.


Structuring Hydrogel Cross-Link Density Using Hierarchical Filament 3D Printing.
Polymer hydrogels, water-laden 3D cross-linked networks, find broad application as advanced biomaterials and functional materials because of their biocompatibility, stimuli responsiveness, and affordability. The cross-linking density reports material properties such as elasticity, permeability, and swelling propensity. However, this critical design parameter can be challenging to template locally. Here, we report a continuous processing scheme that uses laminar flow to direct the organization of cross-linking density across a single sample. Dilute and concentrated poly(ethylene glycol) diacrylate solutions are fed into custom serpentine millifluidic devices. These feature a modular sequence of splitting, rotation, and recombination elements, which create patterned streamlines that serve as a template for hierarchical concentration distributions. Poly(acrylic acid) microgels impart viscoplasticity, which stabilizes layered flow during multiplication and ensures reliable advection. The devices produce structured, seamless filaments, which are then arranged into objects using 3D printing, and photopolymerized to secure the heterogeneous distribution. The flow-encoded, multiscale architecture provides mechanical contrast, which is demonstratively exploited to program robust and reversible shape transformations, potentially useful in soft actuator and sensor applications. The unique structures achieved, and the geometrically dictated, chemistry-agnostic operating principles used to achieve them, provides a new means to engineer hydrogels to suit a variety of applications.


A Rare Case of Transverse Sinus Venous Thrombosis Simulating Postdural Puncture Headache After Cervical Epidural Injection.
Postdural puncture headache (PDPH) is a feared complication related to epidural steroid injections. We report a unique case in which all subjective and objective findings indicated the diagnosis of PDPH. However, the patient failed appropriate conservative and interventional management. Therapeutic failure prompted further investigation to establish the correct diagnosis of cerebral venous sinus thrombosis. Cerebral venous sinus thrombosis is a rare, difficult to diagnose, but potentially lethal disorder with nonspecific and variable clinical presentations, including headache and focal neurological deficits. Performing magnetic resonance imaging and magnetic resonance venogram should be considered early, especially in patients who fail to respond to standard interventions for PDPH.


Assessing the influence of non-treatment variables in a study of outcome from severe head injuries.
An evaluation was made of the impact of non-treatment variables on severely injured head trauma patients. The principal findings were: 1) severity of injury was the best predictor of outcome; 2) patient's age had a statistically significant but marginally useful impact on outcome; 3) a regression analysis showed that duration of transport, up to 4 hours, had no impact on outcome; 4) time from accident to intubation had a marginal impact on outcome; and 5) one-way analysis of variance showed that mode of transportation, whether helicopter, ambulance, or other means had no impact on outcome.


Morphological evidence for uncontrolled proteolytic activity during the ovulatory process in indomethacin-treated rats.
The ovulatory process in cyclic rats was studied after prostanoid synthesis was blocked using indomethacin. Animals were injected at 12:00 h in pro-oestrus with 1.0 mg indomethacin or vehicle (olive oil) and killed at 18:30 h in pro-oestrus, at 02:00, 09:00 and 19:00 h in oestrus and at 09:00 h in metoestrus. Additional rats injected with 0.5 or 4.0 mg indomethacin were killed at 09:00 h in oestrus. No differences in either morphology or serum LH concentrations were found between vehicle or indomethacin-treated rats at 18:30 h in pro-oestrus. However, from 02:00 h in oestrus onward, the process of follicle rupture was altered considerably in indomethacin-treated rats, irrespective of the dose. Early vascular changes, observed in control rats at the apex of the follicle, were absent in indomethacin-treated rats. In some follicles, disruption of the theca layers, invasion of the perifollicular tissue by granulosa cells and follicular fluid, and release of the oocyte to the ovarian interstitium were observed at 02:00 h in oestrus. A small number of follicles ruptured at the ovarian surface. Furthermore, invasion of interstitial tissue, rupture of blood vessel walls, production of emboli of granulosa cells and follicular fluid, and inflammatory reactions were observed in oestrus and metoestrus. The results of the present study demonstrate uncontrolled proteolytic activity, and indicate that abnormal follicle rupture (but not inhibition of follicle rupture) is responsible for ovulation failure in indomethacin-treated rats.


Protective effect of Peucedanum praeruptorum Dunn extract on oxidative damage of LLC‑PK1 cells induced by H2O2.
Peucedanum praeruptorum Dunn extract (PPDE) is a well-known treatment used in traditional Chinese medicines, where it is most commonly used to treat coughs and symptoms such as headaches and fever. In the present study, the antioxidant capacity of PPDE in vitro was determined by scavenging experiments using DPPH, ABTS+·, ·OH, and ·O2-. The cell survival rate was determined by MTT assay. The MDA, SOD, CAT, GSH, and GSH-Px content were determined by colorimetry assays. The expression levels of antioxidant genes SOD, CAT, GSH, and GSH-Px were assessed by reverse transcription-quantitative PCR. HPLC was used to identify the PPDE components. The results suggested that PPDE had scavenging effects on DPPH, ABTS, hydroxyl, and superoxide anion radicals in a concentration-dependent manner; H2O2 treatment resulted in oxidative stress in LLC-PK1 cells, and the degree of injury of LLC-PK1 cells following PPDE treatment was improved, which was positively correlated with its concentration. Peucedanum praeruptorum Dunn extract treatment reduced the content of MDA and increased the content of CAT, SOD1, GSH, and GSH-Px. The mRNA expression levels of antioxidant genes detected by quantitative PCR were consistent with changes in CAT, SOD, GSS, and GSH-Px. Additionally, the trend in CAT, SOD1, GSH, and GSS protein expression levels was also consistent at the mRNA level. PPDE was found to consist of isochlorogenic acid C, myricetin, baicalin, luteolin, and kaempferol. Therefore, PPDE, which was formed of products derived from natural substances, functioned in the inhibition of oxidative damage. The present study aimed to obtain a better understanding of the traditional Chinese medicine Peucedanum praeruptorum Dunn and preliminarily elucidate its antioxidant mechanism at the cellular level. Further animal or human experiments are required to verify the antioxidant effects of PPDE for further development and utilization.


The dependence of human decidual prolactin production and secretion on the osmotic environment in vitro.
Human decidual tissue from uncomplicated term pregnancies was incubated in vitro using a cross-over design of incubation lasting for 72 h. The decidual issue of each membrane was added sequentially to media with the osmolalities 252, 315 and 387 mmol/kg, and the different osmolalities were in 33 experiments induced by changing the concentration either of sodium chloride, potassium chloride, mannitol, sucrose, or choline chloride. At 387 mmol/kg all substances elicited a significant increase in Prl secretion compared with the 315 mmol/kg media (14-27%) or the 252 mmol/kg media (26-46%). When the sodium chloride, potassium chloride, sucrose, and choline chloride media at 387 mmol/kg were compared in another 7 experiments, potassium chloride increased Prl secretion more effectively than the others (10%, P less than 0.05). The incubation cross-over design did not permit determination of the decidual Prl content after incubation in each of the various media, but in 28 experiments the mean decidual Prl content was 12.7 mIU per 100 mg dry tissue (range 4-30) before and 8.7 mIU per 100 mg dry tissue (1-17) after the experiments. The mean amount of Prl secreted during the successive incubations was 152.2 mIU per 100 mg dry tissue (19-672) which showed the secretion to be a result of continued Prl production. The effect of osmolality was independent of the basal production rate. These results indicate that the intracellular ionic concentrations, probably of potassium ion or of chloride ion, are of importance in the regulation of the synthesis and secretion of decidual Prl in vitro.


Infrapatellar Fat Pad-Derived Stem Cell-Based Regenerative Strategies in Orthopedic Surgery.
Infrapatellar fat pad is a densely vascularized and innervated extrasynovial tissue that fills the anterior knee compartment. It plays a role in knee biomechanics as well as constitutes a source of stem cells for regeneration after knee injury. Infrapatellar fat pad-derived stem cells (IPFP-ASCs) possess enhanced and age-independent differentiation capacity as compared to other stem cells, which makes them a very promising candidate in stem cell-based regenerative therapy. The aims of this review are to outline the latest advances and potential trends in using IPFP-ASCs and to emphasize the advantages over other sources of stem cells for applications in orthopedic surgery.


Angiotensin II--nitric oxide interactions in the control of sympathetic outflow in heart failure.
Activation of the sympathetic nervous system is a compensatory mechanism which initially provides support for the circulation in the face of a falling cardiac output. It has been recognized for some time that chronic elevation of sympathetic outflow with the consequent increase in plasma norepinephrine, is counterproductive to improving cardiac function. Indeed, therapeutic targeting to block excessive sympathetic activation in heart failure is becoming a more accepted modality. The mechanism(s) by which sympathetic excitation occurs in the heart failure state are not completely understood. Components of abnormal cardiovascular reflex regulation most likely contribute to this sympatho-excitation. However, central mechanisms which relate to the elaboration of angiotensin II (Ang II) and nitric oxide (NO) may also play an important role. Ang II has been shown to be a sympatho-excitatory peptide in the central nervous system while NO is sympatho-inhibitory. Recent studies have demonstrated that blockade of Ang II receptors of the AT(1) subtype augments arterial baroreflex control of sympathetic nerve activity in the heart failure state, thereby predisposing to a reduction in sympathetic tone. Ang II and NO interact to regulate sympathetic outflow. Blockade of NO production in normal conscious rabbits was only capable of increasing sympathetic outflow when accompanied by a background infusion of Ang II. Conversely, providing a source of NO to rabbits with heart failure reduced sympathetic nerve activity when accompanied by blockade of AT(1) receptors. Chronic heart failure is also associated with a decrease in NO synthesis in the brain as indicated by a reduction in the mRNA for the neuronal isoform (nNOS). Chronic blockade of Ang II receptors can up regulate nNOS expression. In addition, exercise training of rabbits with developing heart failure has been shown to reduce sympathetic tone, decrease plasma Ang II, improve arterial baroreflex function and increase nNOS expression in the central nervous system. This review summarizes a large number of studies which have concentrated on the mechanisms of sympatho-excitation in heart failure. It now seems clear that one mechanism which is important in regulating sympathetic outflow in this disease state depends upon a central interaction between Ang II and NO at the cellular and nuclear levels.


Hyper-regulation of pyr gene expression in Escherichia coli cells with slow ribosomes. Evidence for RNA polymerase pausing in vivo?
UTP-modulated attenuation of transcription is involved in regulating the synthesis of pyrimidine nucleotides in Escherichia coli. Thus, expression of two genes, pyrBI and pyrE, was shown to be under this type of control. The genes encode the two subunits of aspartate transcarbamylase and orotate phosphoribosyltransferase respectively. The levels of these enzymes are inversely correlated with the intracellular concentration of UTP. Modulation of attenuation seems to be a consequence of the effect of UTP concentration on the mRNA chain growth rate. Reducing the UTP pool retards RNA polymerase movement. Mechanistically this will couple the ribosomes translating a leader peptide gene more tightly to the elongating RNA polymerase. The ribosomes will then be more prone to prevent the folding of the mRNA chains into terminating hairpin structures when RNA polymerase is at the attenuator and has to decide whether transcription should terminate or continue into the structural genes. This paper described a study of pyrBI and pyrE gene regulation in cells where the ribosomes move slowly as a result of mutation in rpsL. It appears that expression of the two genes is hyper-regulated by the UTP pool in this type of cells. Furthermore, the attenuator model can only account for the results if it is assumed that UTP-concentration-dependent pausing of transcription occurs in vivo in the two pyr gene leaders such that RNA polymerase waits for the coupled ribosomes before transcribing into the attenuator regions.


Usefulness of the simultaneous bioassay analysis method used for an intake estimation of a radionuclide.
An intake of a radionuclide is estimated based on bioassay measurement data obtained by an in vivo or an in vitro method. Often the intake estimates from one bioassay analysis are considerably different from other results. For better estimates, a simultaneous or combined analysis of measurement data from different bioassay methods is attempted. In this study, the usefulness of a simultaneous bioassay analysis was investigated by using the IDEAS/IAEA intercomparison exercise data and the Individual Monitoring of an Internal Exposure computer code. Tests were made for whole-body counting and urine assay against an acute inhalation of types M and S 60Co particles with various activity median aerodynamic diameter (AMAD). The data set excluding rogue data as well as all the available data were used in this study. The best estimated intake was evaluated based on the best-fit time of an intake determined by minimizing the mean relative deviation Dr. In the case of the whole-body and urine bioassay by using the data excluding some rogue data, the smallest Dr appears at 0.1 and 10 microm of AMAD, respectively, which are different from those estimated by using all the available data. In the case of the simultaneous analysis, it appears at 20 microm of AMAD, which is the same as that estimated by using all the available data. Supposing that monitoring data of a good quality is available, it is expected that the application of a simultaneous analysis to different bioassay methods can provide not only better estimates of an intake but also insights into the validity of the models and parameters used in an interpretation.


Non-invasive imaging of disrupted protein homeostasis induced by proteasome inhibitor treatment using chemical exchange saturation transfer MRI.
Proteasome inhibitors (PIs) are now standard of care for several cancers, and noninvasive biomarkers of treatment response are critically required for early patient stratification and treatment personalization. The present study evaluated whether chemical exchange (CEST) magnetic resonance imaging (MRI) can provide measurements that can be used as the noninvasive biomarkers of proteasome inhibition, alongside diffusion MRI and relaxometry. The sensitivity of human colorectal carcinoma cells to the PI Ixazomib was assessed via in vitro and in vivo dose-response experiments. Acute in vivo response to Ixazomib was assessed at three dosing concentrations, using CEST MRI (amide, amine, hydroxyl signals), diffusion MRI (ADC) and relaxometry (T1, T2). These responses were further evaluated with the known histological markers for Ixazomib and Bradford assay ex vivo. The CEST signal from amides and amines increased in proportion to Ixazomib dose in colorectal cancer xenografts. The cell lines differed in their sensitivity to Ixazomib, which was reflected in the MRI measurements. A mild stimulation in tumor growth was observed at low Ixazomib doses. Our results identify CEST MRI as a promising method for safely and noninvasively monitoring disrupted tumor protein homeostasis induced by proteasome inhibitor treatment, and for stratifying sensitivity between tumor types.


[Effect on the recovery process of nasal sinus mucosa with nasosinusitis and nasal polyps influenced by applying Chinese herbal preparation].
To observe the action of Chinese herbal preparation (CHP) on the recovery of nasal sinus mucosa after endoscopic sinus surgery (ESS). Seventy-eight patients (140 sinus) with chronic nasosinusitis and nasal polyps, who had undergone ESS, were divided into two groups. The 40 patients (70 sinuses) in the experiment group were treated with daily nasal flushing with diluted CHP while the 38 patients (70 sinuses) in the control group were untreated. Tissues of nasal mucosa were taken out from patients' posterior walls of maxillary sinus at different time points, i. e. 2 - 3 weeks, 8 - 11 weeks and 13 - 15 weeks after ESS were observed. Significant difference was shown between the two groups under endoscope at all the time points in occurrence (patient/sinus) of sticky mucus, swelling and thickened mucus, occlusion of sinus opening, bloody secretion in sinus, and adhesion, etc. (P < 0.05, P < 0.01). Light microscopic examination showed significant difference between the two groups in occurrence (patient/sinus) of squamous epithelial metaplasia and fiber tissues proliferation (P <0.05). And electron microscopic examination also showed significant difference between the two groups in occurrence (patient/sinus) of inflammatory cell infiltration, fibers decrease, disordered arrangement, microvilli and short cilia (P<0.01). Using CHP for postoperative nasal flushing after ESS is a safe and manageable approach with effects in promoting nasal mucosa recovery.


Contrast adaptation and contrast masking in human vision.
After a preliminary study of visual evoked potentials (VEPS) to a test grating seen in the presence of masks at different orientations, psychophysical data are presented showing the effects of adaptation and of masking on thresholds for detecting the same test grating. The test is a vertical grating of spatial frequency 2 cycles per degree; adapting and masking gratings differ from the test either in orientation or in spatial frequency. The effects of adaptation and masking are explained by a single mechanism model that assumes: (i) adaptation and masking both alter the contrast response (or transducer) function of the mechanism that detects the test; (ii) masks, but not adaptors, stimulate the mechanism that detects the test; and (iii) a test is detectable when it raises response level by a constant amount. The model incorporates two distinct tuning functions, a broad adaptive contrast function and a narrow effective contrast function. It accounts adequately for all the data, including the location and size of the facilitative dip found in some masking functions, the constant slopes of the threshold elevation segments of adaptation functions and the varying slopes of masking functions. It also predicts the sometimes surprising joint effects of adaptation followed by masking and of two masks operating simultaneously.


Stabilization of fully reduced iron-sulfur clusters by carbene ligation: the [FenSn]0 oxidation levels (n = 4, 8).
The all-ferrous [Fe4S4](0) state has been demonstrated in the fully reduced Fe protein of the Azotobacter vinelandii nitrogenase complex. We seek synthetic analogues of this state more tractable than the recently prepared but highly unstable cluster [Fe4S4(CN)4](4-) (Scott, Berlinguette, Holm, and Zhou, Proc. Natl. Acad. Sci. U.S.A. 2005, 102, 9741). The N-heterocyclic carbene 1,3-diisopropyl-4,5-dimethylimidazol-2-ylidene (Pr(i)2NHCMe2) has been found to stabilize the fully reduced clusters [Fe8S8(Pr(i)2NHCMe2)6] (4) and [Fe4S4(Pr(i)2NHCMe2)4] (5), which are prepared by cluster assembly or phosphine substitution of FenSn (n = 8, 16) clusters. Cluster 4 is also obtained by reaction of the carbene with all-ferrous [Fe7S6(PEt3)5Cl2] (3) and cluster 5 by carbene cleavage of 4. Detailed structures of 3 (monocapped prismatic), 4, and 5 are described; the latter two are the first iron-sulfur clusters with Fe-C sigma bonds. Cluster 4 possesses the [Fe8(mu3-S) 6(mu4-S)2] edge-bridged double cubane structure and 5 the cubane-type [Fe4(mu3-S)4] stereochemistry. The all-ferrous formulations of the clusters are confirmed by X-ray structure parameters and (57)Fe isomer shifts. Both clusters are stable under conventional aprotic anaerobic conditions, enabling further study of reactivity. The collective properties of 5 indicate that it is a meaningful synthetic analogue of the core of the fully reduced protein-bound cluster.


Bandwidth-tunable single-photon source in an ion-trap quantum network.
We report a tunable single-photon source based on a single trapped ion. Employing spontaneous Raman scattering and in-vacuum optics with large numerical aperture, single photons are efficiently created with controlled temporal shape and coherence time. These can be varied between 70 ns and 1.6 micros, as characterized by operating two sources simultaneously in two remote ion traps which reveals mutual and individual coherence through two-photon interference.


Toward program development.
An era of disciplined thinking may result not only in acknowledging but also in addressing those questions that may lead to a consistent examination of existing programs within a profession. This paper is one attemp at encouraging the professional membership to continue to address: 1. the nature and focus of the content and practice environments of the field of Occupational Therapy; 2. two approaches to investigating the distinctive character of the field of Occupational Therapy-the Separate Domain Approach and the Retroductive Approach; 3. and the basic documents developed and/or sponsored by the professional organization-in this case-The Ohio State Study (1), the NIH Study (2), and the Curriculum Guide (3).


Optical control of thermocapillary effects in complex nanofluids.
We study the strong coupling of light and nanoparticle suspensions and their surface tension effect in capillaries. We show experimentally and theoretically that increasing the intensity of a narrow laser beam passing through a capillary far away from the surface results in a significant decrease in the fluid level. The underlying mechanism relies on light-induced redistribution of nanoparticles in the bulk and the surface of the fluid, facilitating continuous optical control over the surface position. The experiments manifest optical control from afar over properties of fluid surfaces.


Acridine-modified, clamp-forming antisense oligonucleotides synergize with cisplatin to inhibit c-Myc expression and B16-F0 tumor progression.
The c-myc protooncogene plays a key role in the abnormal growth regulation of melanoma cells. We have targeted three polypurine sequences within the mouse myc mRNA with acridine-modified, clamp-forming antisense oligonucleotides (AS ODNs) in an effort to inhibit growth of murine melanoma cells. These ODNs are unique in that they hybridize to the target mRNA by both Watson-Crick and Hoogsteen hydrogen bond interactions, forming a triple-stranded structure. At a concentration of 3 microM E1C, E2C and E3C inhibit B16-F0 proliferation by 76, 66 and 78%, respectively. Both immunofluorescent staining and western blot analysis corroborate a proportional reduction in c-Myc expression by all three ODNs. There were clear distinctions in the ability of these ODNs to inhibit tumor progression in C57BL/6 mice as a function of Myc expression. There was no synergy demonstrated between ODN E1C with cisplatin (DDP), which inhibited tumor growth by 77% alone and 82% in combination. Although E2C inhibited growth by 54%, its effect was decreased to 32% with DDP, when compared with controls. E3C, on the other hand, demonstrated a synergistic effect with DDP, inhibiting growth by 72% in combination, but only by 1% as a single agent. Immunofluorescence analysis of tumors for each group revealed a concomitant reduction in c-Myc expression in tumors from mice treated with the most active clamp ODN alone (E1C) or clamp ODN + DDP (E1C/E3C + DDP). Western blot analysis confirmed this decrease in target protein expression. Our results document the growth-inhibitory activity of two myc-targeting antisense clamp ODNs; E1C, which has activity as a single agent, and E3C, which has in vivo synergy with DDP pretreatment. These data confirm the antiproliferative effects of these novel ODNs and document an interesting synergy with the chemotherapeutic agent DDP.


Time-resolved fluorescent X-ray interference.
A fluorescent X-ray interference method can effectively measure nanometer-level conformational changes for non-crystallized molecules and proteins in aqueous conditions. The time-resolved technique can be used to obtain information about the dynamics of molecules and proteins. Instrumentation for time-resolved fluorescent X-ray interference has been designed. A typical interference-fringe pattern was observed with approximately 3 s of X-ray exposure time from K-fluorescent X-rays emitted from a Zn monoatomic layer on an Rh substrate. The primary X-ray beam was polychromed with a mirror for total external reflection of X-rays and was tuned to an energy level at which only Zn K radiation became optimally excited. The glancing angle of the primary X-ray beam was fixed at a glancing angle at which the total intensity of K-fluorescent X-rays emitted from Zn atoms corresponded to the maximum value. The fluorescent X-ray interference fringes were monitored with an imaging plate (IP) as a non-energy-dispersive two-dimensional detector. The exposed interference fringes on the IP were integrated along the direction of the fringes. The integrated fringes were in close agreement with a theoretical estimate based on the interference among transmitted and reflected waves at interfaces in the sample.


Phase III, Randomized Study of Atezolizumab Plus Bevacizumab and Chemotherapy in Patients With EGFR- or ALK-Mutated Non-Small-Cell Lung Cancer (ATTLAS, KCSG-LU19-04).
In the treatment of non-small-cell lung cancer (NSCLC) with a driver mutation, the role of anti-PD-(L)1 antibody after tyrosine kinase inhibitor (TKI) remains unclear. This randomized, open-label, multicenter, phase III study evaluates the efficacy of atezolizumab plus bevacizumab, paclitaxel, and carboplatin (ABCP ) in EGFR- or ALK-mutated NSCLC that progressed before TKI therapy. We compared the clinical efficacy of ABCP followed by maintenance therapy with atezolizumab plus bevacizumab with pemetrexed plus carboplatin or cisplatin (PC) followed by pemetrexed maintenance. The primary end point was progression-free survival (PFS). A total of 228 patients with activating EGFR mutation (n = 215) or ALK translocation (n = 13) were enrolled from 16 sites in the Republic of Korea and randomly assigned at 2:1 ratio to either ABCP (n = 154) or PC arm (n = 74). The median follow-up duration was 26.1 months (95% CI, 24.7 to 28.2). Objective response rates (69.5% v 41.9%, P < .001) and median PFS (8.48 v 5.62 months, hazard ratio [HR], 0.62 [95% CI, 0.45 to 0.86]; P = .004) were significantly better in the ABCP than PC arm. PFS benefit increased as PD-L1 expression increased, with an HR of 0.47, 0.41, and 0.24 for PD-L1 ≥1%, ≥10%, and ≥50%, respectively. Overall survival was similar between ABCP and PC arm (20.63 v 20.27 months, HR, 1.01 [95% CI, 0.69 to 1.46]; P = .975). The safety profile of the ABCP arm was comparable with that previously reported, with no additional safety signals, but higher rates of treatment-related adverse events were observed compared with the PC arm. To our knowledge, this study is the first randomized phase III study to demonstrate the clinical benefit of anti-PD-L1 antibody in combination with bevacizumab and chemotherapy in patients with EGFR- or ALK-mutated NSCLC who have progressed on relevant targeted therapy.


Controlled release and reversal of multidrug resistance by co-encapsulation of paclitaxel and verapamil in solid lipid nanoparticles.
Paclitaxel (PTX) has been used in the treatment of wide range of cancers but its entry into cancer cell is restricted by p-glycoprotein (p-gp). Also, it was reported that verapamil (VP) could inhibit p-gp efflux. Hence, three kinds of solid lipid nanoparticles (SLN) such as PVS (PTX and VP co-loaded SLN), PSV (PTX loaded SLN, later added VP) and PVSV (PTX and VP co-loaded SLN, later added VP) were prepared to overcome MDR by combination of PTX and VP. PVS was the SLN loaded with both PTX and VP at the same time. PSV was the SLN loaded with PTX and then modified with VP - complexed hydroxypropyl-β-cylcodextrin (HPCD). Finally, PVSV was the SLN loaded with PTX and half of VP at the same time subsequently, modified with half of VP - complexed HPCD. The physicochemical characterizations of PVS, PSV or PVSV such as particle size, zeta potential, encapsulation efficiency or in vitro PTX release were examined. PVSV showed that release of VP was higher than PTX solution in first 15h and sustained release of both VP and PTX. PVSV showed significantly higher cytotoxicity and cellular uptake than that of the PTX solution in MCF-7/ADR resistant cells. Furthermore, PVSV significantly down regulated the expression of p-gp than the PTX solution in MCF-7/ADR resistant cells. Based on these findings, this study indicated that the PVSV exhibited great potential for breast cancer therapy.


Effects of enhanced metal recovery on the recycling potential of MSWI bottom ash fractions in various legal frameworks.
In recent years, complex new bottom ash treatment processes for enhanced metal recovery have been implemented in Switzerland, producing residual bottom ash fractions with various qualities. This study focusses on three different treatment processes by characterizing all arising fractions in detail. Thereby the factors influencing the composition of these fractions are identified and their recycling potential in Switzerland is investigated. However, high legislative requirements on total contents of heavy metals represent a high barrier for bottom ash recycling in Switzerland. Therefore, the recycling potential is further evaluated based on the waste legislation applied in the Netherlands, where recycling of bottom ash has a long tradition. There, threshold values for bottom ash recycling are based on leachate concentrations and not on total contents as in Switzerland. However, Swiss Waste Legislation also knows threshold values based on leachate concentrations for certain waste materials. The leaching tests applied in these two countries, however, are different. The comparison of both leaching tests reveals that the setup and conditions, especially the considered pH range, significantly influence the leaching of heavy metals. With emphasis on problematic pollutants, the possibilities for new applications of these fractions are evaluated based on Swiss and Dutch legal threshold values. The comparison within the legal frameworks of these two countries allows recognizing opportunities and risks related to bottom ash recycling.


Transosseous Fixation of the Distal Tibiofibular Syndesmosis: Comparison of Interosseous Suture and Endobutton Across Age Groups.
Background In the ankle, suture bridge fixation for syndesmotic injuries is commonly employed. Initial recommendations for suture bridge constructs advised against using the device in patients with insufficient quantity or quality of bone. Therefore, many surgeons limit its use to younger, more athletic patients and use traditional screw fixation in older, less active patients. The purpose of this study is to compare the outcomes of suture bridge fixation for syndesmotic repair in patients ≥ 60 years old vs patients < 60 years old. Methods A retrospective review of 140 ankle fracture patients from a single institution who received suture bridge fixation between July 13, 2010, and February 2, 2022, was performed. Patient data was obtained from patient records in the electronic health record. Univariate analysis, including chi-square and independent t-tests, was used. Complications included delayed wound healing, infection, hardware loosening, and non-union. Results There were no significant differences in demographics, comorbidities, primary or other procedures, loss of fixation, and neuropathy between groups. There was also no difference within the distribution of the mechanism of injury, affected side, or Weber classification. Finally, the rate of complication and complication type showed no significant differences between patients 60 years and older versus 60 years and younger. Complication rates and types in patients > 60 years versus < 60 years were not significantly different. Conclusion The use of the suture bridge fixation in patients > 60 years may not lead to an increased risk of complications and appears to be safe for use.


Quantification of cells recovered by bronchoalveolar lavage. Comparison of cytocentrifuge preparations with the filter method.
Controversy exists as to the appropriate methods to use in the processing of bronchoalveolar lavage (BAL) fluid for total cell numbers and cellular differential analysis. It has been shown that cell losses (primarily lymphocytes) occur by the most commonly employed methods. Therefore, we examined the total cell and differential counts obtained by several methods of cytocentrifuge preparation and by the filter preparation in 46 consecutive patients with interstitial lung disease and 29 healthy volunteers undergoing bronchoalveolar lavage. The retrieved lavage fluid was pooled, and an aliquot was used to determine the total cell count, cell viability, and the differential cell count by the filter and cytocentrifuge techniques. The remaining fluid was centrifuged (800 g for 10 min), and the cell pellet was resuspended in Hank's balanced salt solution without Ca2+ and Mg2+. An aliquot of these centrifuged and resuspended cells was used for repeat determination of the cell viability, total cell count, and cellular differential by cytocentrifuge technique. Autologous serum was added to another aliquot of these centrifuged and resuspended cells to arrive at a 10% protein solution, and the cellular differential obtained by cytocentrifuged preparation.(ABSTRACT TRUNCATED AT 250 WORDS)


Validation of triggers and development of a pediatric trigger tool to identify adverse events.
Little is known about adverse events (AEs) in pediatric patients. Record review is a common methodology for identifying AEs, but in pediatrics the record review tools generally have limited focus. The aim of the present study was to develop a broadly applicable record review tool to identify AEs in pediatric inpatients. Using a broad literature review and expert opinion with a modified Delphi process, a pediatric trigger tool with 88 triggers, definitions, and descriptions including AE preventability decision support was developed and tested in a random sample of 600 hospitalized pediatric patients admitted in 2010 to a single university children's hospital. Four registered nurse-physician teams performed complete two-stage retrospective reviews of 150 records each from either neonatal, surgical/orthopedic, medicine, or emergency medicine units. Registered nurse review identified 296 of 600 records with triggers indicating potential AEs. Records (n = 121) with only false positive triggers not indicating any potential AEs were not forwarded to the next review stage. On subsequent physician review, 204 (34.0%) of patients were found to have had 563 AEs, range 1-27 AEs/patient. A total of 442 preventable AEs were found in 161 patients (26.8%), range 1-22. Overall, triggers were found 3,598 times in 417 (69.5%) records, with a mean of 6 (median 1, range 0-176) triggers per patient. The overall positive predictive value of the triggers was 22.9%, (range 0.0-100.0%). The final pediatric trigger tool, developed with a second Delphi round, required 29 triggers. AEs are common in pediatric patients and most are preventable. The main contributions of this study are to further develop and adapt trigger definitions, including AE preventability decision support, to introduce new triggers in pediatric care, as well as to apply pediatric triggers in different clinical specialties. Our findings resulted in a national pediatric trigger tool, and might also be adapted internationally. The pediatric trigger tool can help healthcare organizations to measure and analyze the AEs occurring in hospitalized children in order to improve patient safety.


Human chorionic gonadotropin β regulates epithelial-mesenchymal transition and metastasis in human ovarian cancer.
Human chorionic gonadotropin β (β-hCG) is a well-known and accurate marker for the diagnosis and monitoring of pregnancy, trophoblastic tumors and ovarian germ cell tumors. Recently, β-hCG has been found to be closely related to poor prognosis and metastasis in various other malignant tumors, while its role and mechanism in ovarian cancer is still unclear. In the present study, lentiviral‑mediated transfection and small interfering RNA (siRNA) were used to alter β-hCG expression in the ovarian cancer cell lines ES-2 and SKOV3, respectively. Then, migration and invasion activity regulated by β-hCG were evaluated by wound-healing and Transwell assays in vitro and in a peritoneal xenograft nude mouse model in vivo. EDTA and trypsin were utilized to investigate the attachment ability of these cells. Moreover, the expression of epithelial mesenchymal transition (EMT) markers (β-catenin, Slug, vimentin, Snail, claudin, E-cadherin and N-cadherin) was assessed by western blotting and immunofluorescence in ES-2 and SKOV3 cells. Furthermore, β-hCG and EMT markers were evaluated in human ovarian cancer specimens by IHC. The results showed that overexpression of β-hCG clearly promoted migration and invasion in ES-2 and SKOV3 cells (P<0.05) and facilitated metastasis in peritoneal xenografts, while silencing of β-hCG led to the opposite effect. Moreover, β-hCG was closely associated with cell morphology, attachment ability and EMT marker expression in ES-2 and SKOV3 cells and human ovarian cancer specimens. Upregulation of β-hCG promoted cells from an epithelial-like morphology to a mesenchymal-like phenotype, decreased the adhesion ability (P<0.05), and reduced the expression of epithelial markers (E-cadherin) while inducing the expression of mesenchymal markers (vimentin, N-cadherin, β-catenin and Slug). Furthermore, the converse effects were confirmed by knockdown of β-hCG. These findings strongly suggest that β-hCG may regulate metastasis of ovarian cancer through EMT, and it may become a new target for therapeutic intervention.


A microbiological study of bottled mineral water marketed in Ludhiana.
The microbiological quality of bottled mineral water marketed in Ludhiana was examined, Twenty three brands were analyzed for presumptive coliform count by multiple tube tests, and E. coli count was confirmed by Eijkman test. Bacterial and fungal loads were tested by membrane filtration test. Out of 23 only one sample (4.4%) showed the presumptive coliform count to be 460 most probable number (MPN)l 1 00ml,and 1 was found to be positive when tested by Eijkman test for Ecoli. In the membrane filtration test three samples (13%) showed more than two types of bacteria. Different types of bacteria isolated included Bacillus sp (19/23). Pseudomonas spp (13123), Ecoli, Klebsiella sp and S.albus one each Fungi was isolated from five of twenty three. (22%) samples. Only one brand of mineral water was unfit for human consumption. The rest of the samples were contaminated with non pathogenic flora.


Contribution of nerve biopsy to unclassified neuropathy.
To evaluate the diagnostic yield of nerve biopsy in patients with peripheral neuropathy of undetermined cause despite extensive diagnostic workup. From November 2001 through January 2004, 38 patients underwent nerve biopsy because of unclassified neuropathy. The etiology of the neuropathies could be defined in 14 patients (37%), i.e. in 15% of chronic symmetric, 30% of chronic asymmetric, 50% of subacute symmetric and 62.5% of subacute asymmetric neuropathies. The biopsy was diagnostic in 6 patients (16%), where it showed a vasculitis, and supportive in 8 patients (21%). The contribution of nerve biopsy to the diagnosis of peripheral neuropathy was highest in acute and subacute asymmetric forms of neuropathy and lowest in chronic symmetric forms. The main indication for nerve biopsy remains the diagnosis of vasculitic neuropathy, a potentially treatable disorder.


The Color of Cation-π Interactions: Subtleties of Amine-Tryptophan Interaction Energetics Allow for Radical-like Visible Absorbance and Fluorescence.
Several peptides and a protein with an inter- or intramolecular cation-π interaction between tryptophan (Trp) and an amine cation are shown to absorb and fluoresce in the visible region of the spectrum. Titration of indole with sodium hydroxide or ammonium hydroxide yields an increasing visible fluorescence as well. Visible absorption and multipeaked fluorescence excitation spectra correlate with experimental absorption spectra and the vibrational modes of calculated absorption spectra for the neutral Trp radical. The radical character of the cation-indole interaction is predicted to stem from the electrostatic dislocation of indole highest occupied molecular orbital (HOMO) charge density toward the cation with a subsequent electronic transition from the HOMO-2 to the HOMO. Because this is a vertical transition, fluorescence is possible. Hydrogen bonding at the indole amine most likely stabilizes the radical-like state. These results provide new spectroscopic tools for the investigation of cation-π interactions in numerous biological systems, among them, proteins and their myriad ligands, and show that one, or at most, two, point mutations with natural amino acids are all that is required to impart visible fluorescence to proteins.


Rapid development of hepatic tumors in transforming growth factor alpha transgenic mice associated with increased cell proliferation in precancerous hepatocellular lesions initiated by N-nitrosodiethylamine and promoted by phenobarbital.
The carcinogenic and tumor-promoting effects of human transforming growth factor alpha (TGF-alpha) overexpression were examined in a two-stage chemical carcinogenesis protocol using TGF-alpha transgenic mouse line MT42. Male MT42 and CD-1 mice received a single i.p. injection of 5 mg N-nitrosodiethylamine (DEN)/kg body wt at 15 days of age, and were placed on a diet containing 0.05% of phenobarbital (PB) from 4 weeks of age for 35 weeks. DEN-, PB-treated and saline-injected animals in each strain were used as controls. A total of three sequential sacrifices (at 10, 23 and 37 experimental weeks) was performed. Hepatocellular carcinomas (HCCs) developed earlier at high incidence (100%) after 23 experimental weeks in MT42 mice receiving DEN/PB, while CD-1 mice had a 40% incidence of HCCs only after week 37. HCCs also developed in the DEN-initiated MT42 mice at 80% incidence after week 23, but no HCCs were observed in the DEN-initiated CD-1 mice. PB induced preneoplastic foci (67%), adenomas (33%) and HCCs (33%) after 37 weeks in MT42 mice, but no lesions were found in CD-1 mice. Thus, the carcinogenic response to DEN and/or PB was accelerated in the MT42 transgenic mice. Furthermore, PB promotion was observed from week 10 in MT42 mice and week 23 in CD-1 mice. Thus, the promoting effect of PB was also accelerated in the MT42 transgenic mice. Proliferating cell nuclear antigen (PCNA) labeling indices of hepatocellular foci and adenomas in DEN- or DEN/PB-treated MT42 mice were significantly higher than those of CD-1 mice. TGF-alpha expression determined by immunohistochemistry revealed higher levels in these lesions than in hepatocytes of surrounding parenchyma of MT42 transgenic mice. In conclusion, TGF-alpha transgenic mice clearly demonstrated enhanced sensitivity to the development of hepatocellular carcinoma in the DEN initiation and PB promotion regime, possibly through a mechanism of increased hepatocyte proliferation in precancerous lesions (foci and adenomas), driven by high expression of the mitogen TGF-alpha in these lesions.


Tests of the regenerative capacity of tectal efferent axons in the frog, Rana pipiens.
Experiments were designed to determine if neurons of the ranid optic tectum, a major target of the optic nerve, possess the same regenerative potential as optic axons. Normal tectal efferent (TE) projections were reexamined by using the anterograde transport of 3H-proline and autoradiography (n = 18), bulk-filling damaged TE axons with horseradish peroxidase (HRP; n = 18) and anterogradely transporting wheat germ agglutinin-HRP (n = 8) to label TE axons. Results were similar to reports that used degeneration methods (Rubinson: Brain Behav. Evol. 1:529-561, '68; Lazar: Acta. Biol. Hung. 20:171-183, '69). Following a brainstem hemisection just caudal to the nucleus isthmi (1-20 weeks), the ipsilateral descending TE pathway was autoradiographically examined (n = 20). While all other TE projections appeared normal, there was no detectable ipsilateral descending projection beyond the lesion site. Ascending TE axons were cut at the anterior tectal border by hemisecting the left diencephalon (LDH)--a lesion that also cuts optic axons projecting to the left tectum. There was no indication of TE axonal regeneration with the aid of autoradiography or HRP histochemistry 1-30 weeks postlesion (n = 48) even when the medial diencephalon was intentionally left intact (n = 4). However, in all four cases examined, optic axons regenerated following the same LDH where TE axonal regeneration failed (also see Stelzner, Lyon, and Strauss: Anat. Rec. 205:191A-192A, '83). Local effects of LDH should be similar for both the cut optic and cut TE axons. Other factors were tested that may contribute to the lack of TE axonal regeneration. Our results indicate that optic regeneration itself (n = 8), postaxotomy retrograde cell death of TE neurons (n = 6), deafferentation of the tectum of optic axons, and potential sprouting within tectal targets by intact contralateral TE axons (n = 10) are not critical factors aborting TE axonal regeneration. TE axons filled with HRP at chronic periods after LDH (n = 4) terminate anomalously near the LDH border. Many of these endings are similar to reactive endings or terminal clubs seen after axonal injury in the mammalian CNS. Our results suggest that this disparity in regenerative ability of optic and TE axons may be related to a difference in the responsive ability of these cell types to initiate or maintain axonal elongation after axotomy within the amphibian CNS environment.


Combination of natural killer cell-based immunotherapy and irreversible electroporation for the treatment of hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is among the most lethal cancer types despite great advancement in overall survival of the patients over the last decades. Surgical resection or partial hepatectomy has been approved as the curative treatment for early-stage HCC patients however only up to 30% of them are eligible for the procedures. Natural killer (NK) cells are cytotoxic lymphocytes recognized for killing virally infected cells and improving immune functions for defending the body against malignant cells. Although autologous NK cells failed to demonstrate significant clinical benefit, transfer of allogeneic adoptive NK cells arises as a promising approach for the treatment of solid tumors. The immunosuppressive tumor microenvironment and inadequate homing efficiency of NK cells to tumors can inhibit adoptive transfer immunotherapy (ATI) efficacy. However, potential of the NK cells is challenged by the transfection efficiency. The local ablation techniques that employ thermal or chemical energy have been investigated for the destruction of solid tumors for three decades and demonstrated promising benefits for individuals not eligible for surgical resection or partial hepatectomy. Irreversible electroporation (IRE) is one of the most recent minimally invasive ablation methods that destruct the cell within the targeted region through non-thermal energy. IRE destroys the tumor cell membrane by delivering high-frequency electrical energy in short pulses and overcomes tumor immunosuppression. The previous studies demonstrated that IRE can induce immune changes which can facilitate activation of specific immune responses and improve transfection efficiency. In this review paper, we have discussed the mechanism of NK cell immunotherapy and IRE ablation methods for the treatment of HCC patients and the combinatorial benefits of NK cell immunotherapy and IRE ablation.


Remodeling the Epigenetic Landscape of Cancer-Application Potential of Flavonoids in the Prevention and Treatment of Cancer.
Epigenetics, including DNA methylation, histone modification, and noncoding RNA regulation, are physiological regulatory changes that affect gene expression without modifying the DNA sequence. Although epigenetic disorders are considered a sign of cell carcinogenesis and malignant events that affect tumor progression and drug resistance, in view of the reversible nature of epigenetic modifications, clinicians believe that associated mechanisms can be a key target for cancer prevention and treatment. In contrast, epidemiological and preclinical studies indicated that the epigenome is constantly reprogrammed by intake of natural organic compounds and the environment, suggesting the possibility of utilizing natural compounds to influence epigenetics in cancer therapy. Flavonoids, although not synthesized in the human body, can be consumed daily and are common in medicinal plants, vegetables, fruits, and tea. Recently, numerous reports provided evidence for the regulation of cancer epigenetics by flavonoids. Considering their origin in natural and food sources, few side effects, and remarkable biological activity, the epigenetic antitumor effects of flavonoids warrant further investigation. In this article, we summarized and analyzed the multi-dimensional epigenetic effects of all 6 subtypes of flavonoids (including flavonols, flavones, isoflavones, flavanones, flavanols, and anthocyanidin) in different cancer types. Additionally, our report also provides new insights and a promising direction for future research and development of flavonoids in tumor prevention and treatment via epigenetic modification, in order to realize their potential as cancer therapeutic agents.