Complete Penile Necrosis in a Patient With Heparin-induced Thrombocytopenia: A Case Report.
Penile necrosis is a rare condition that has been mostly described in association with diabetes mellitus and end-stage renal disease. We report an unusual case of acute penile necrosis because of heparin-induced thrombocytopenia. A 75-year-old man presented with acute renal failure and experienced cardiac complications during the hospitalization. The patient was treated twice with intravenous heparin. He developed symptoms of penile necrosis 4 days after the reintroduction of heparin. At that moment, the platelet count dropped by 61%, and the analysis of heparin-pf4 antibodies was positive for heparin-induced thrombocytopenia. The patient underwent a total penectomy and a perineal urethrostomy.


Drug dosing and monitoring in obese patients undergoing allogenic stem cell transplantation.
The effects of physiological changes in patients with obesity on pharmacokinetic parameters and the time course of drug response, especially in the field of haematology/oncology, are poorly understood. For some antimicrobial drugs, dosing considerations exist, while for cytostatic drugs, dose modifications for obese patients are not consistently recommended. Glomerular filtration rate and renal perfusion appear to be similar in obese and normal weight individuals, thus elimination of hydrophilic and extensively renally cleared drugs mainly depends upon creatinine clearance. To provide information about drug dosing in morbidly obese patients undergoing allogenic haematopoietic stem cell transplantation and to develop dosing recommendations for those patients, based on literature data, pharmacokinetic properties and own experiences. A review on the literature on drug dosing in obese patients as well as on the pharmacokinetic properties of drugs which are supposed to be used in the field of stem cell transplantation was combined with own data on drug dosing and pharmacokinetic drug monitoring in a morbidly obese patient undergoing matched-unrelated allogenic peripheral blood stem cell transplantation. For hydrophilic and extensively renally cleared drugs (e.g. piperacillin/sulbactam, cotrimoxazole, fludarabine) standard dosages for adult patients or dosing based on ideal body weight (IBW) (e.g. aciclovir, methotrexate) can be used. For ciclosporin and digitoxin we could show that high initial doses are needed to achieve sufficient plasma concentrations. After steady state distribution was completed, maintenance doses comparable to normal weight patients are sufficient. Likewise, distribution of enoxaparin and phenytoin seems to take longer in obese patients. Dosing recommendations of 25 drugs that can be used in morbidly obese patients undergoing allogenic stem cell transplantation are given. Pharmacotherapy in morbidly obese patients undergoing allogenic stem cell transplantation is possible, if pharmacokinetic properties of the drugs are considered and close monitoring of plasma concentrations is performed.


Blinded evaluation of cathepsin S inhibitors from the D3RGC3 dataset using molecular docking and free energy calculations.
During the last few years, we have developed a docking protocol involving two steps: (i) the choice of the most appropriate docking software and parameters for the system of interest using structural and functional information available in public databases (PDB, ChEMBL, PubChem Assay, BindingDB, etc.); (ii) the docking of ligand dataset to provide a prediction for the binding modes and ranking of ligands. We applied this protocol to the D3R Grand Challenge 3 dataset containing cathepsin S (CatS) inhibitors. Considering the size and conformational flexibility of ligands, the docking calculations afforded reasonable overall pose predictions, which are however dependent on the specific nature of each ligand. As expected, the correct ranking of docking poses is still challenging. Post-processing of docking poses with molecular dynamics simulations in explicit solvent provided a significantly better prediction, whereas free energy calculations on a subset of compounds brought no significant improvement in the ranking prediction compared with the direct ranking obtained from the scoring function.


Esophageal resection for cancer.
Currently, relatively safe, reliable resection techniques are available for most patients with esophageal carcinoma who present with nonmetastatic disease. For optimal results, the surgeon must be familiar with both transhiatal and transthoracic approaches and must individualize the approach depending on the tumor size and location and the patient's functional status. Whereas post-resection survival rates are good for patients with early-stage disease (Stage I or IIa), most patients present with locally advanced, Stage III disease. Although some progress has been made in the past decade in regard to early diagnosis among patients with Barrett's metaplasia undergoing endoscopic surveillance and additional progress has been made in adapting multimodality treatment programs successfully to patients with locally advanced disease, the overall cure rate for patients with esophageal carcinoma remains low.


Recombinant production of a diffusible signal factor inhibits Salmonella invasion and animal carriage.
The complex chemical environment of the intestine is defined largely by the metabolic products of the resident microbiota. Enteric pathogens, elegantly evolved to thrive in the gut, use these chemical products as signals to recognize specific niches and to promote their survival and virulence. Our previous work has shown that a specific class of quorum-sensing molecules found within the gut, termed diffusible signal factors (DSF), signals the repression of Salmonella tissue invasion, thus defining a means by which this pathogen recognizes its location and modulates virulence to optimize its survival. Here, we determined whether the recombinant production of a DSF could reduce Salmonella virulence in vitro and in vivo. We found that the most potent repressor of Salmonella invasion, cis-2-hexadecenoic acid (c2-HDA), could be recombinantly produced in E. coli by the addition of a single exogenous gene encoding a fatty acid enoyl-CoA dehydratase/thioesterase and that co-culture of the recombinant strain with Salmonella potently inhibited tissue invasion by repressing Salmonella genes required for this essential virulence function. Using the well characterized E. coli Nissle 1917 strain and a chicken infection model, we found that the recombinant DSF-producing strain could be stably maintained in the large intestine. Further, challenge studies demonstrated that this recombinant organism could significantly reduce Salmonella colonization of the cecum, the site of carriage in this animal species. These findings thus describe a plausible means by which Salmonella virulence may be affected in animals by in situ chemical manipulation of functions essential for colonization and virulence.


Breast cancer dermal lymphatic invasion recurrence and contralateral axillary lymph nodes metastasis after complete response to neoadjuvant therapy: A case report.
Non-luminal type breast cancer patients with pathological complete response (pCR) by neoadjuvant chemotherapy (NAC) usually have a good prognosis, but occasionally recurrence occurs. A 61-year-old woman was diagnosed with breast cancer T2N2aM0 stage IIIA and its intrinsic type was non-luminal type. After NAC, the patient achieved pCR and underwent breast-conserving surgery and axillary lymph node dissection (ALND). Radiotherapy and trastuzumab of one-year duration was added. However, six years and two months later, local recurrence and contralateral axillary lymph node (CLALN) metastasis were identified. After resection, anti-human epidermal growth factor receptor 2 (HER-2) therapy was done, however, six months after operation, purpura was observed on the right chest and tended to increase. One and a half years after re-operation, dermal lymphatic invasion (DLI) recurrence without clinical inflammatory signs was diagnosed. A skin resection was performed >1.5 cm away from the purpura, and the surgical margins were negative but four months later, a recurrence re-emerged. CLALN metastasis is considered distant metastasis based on the current TNM classification. However, as previous ALND or radiotherapy can change lymphatic flow, the resulting CLALN may not be distant metastasis. DLI recurrence without clinical inflammatory signs is likely to be resistant to anti-HER2 even in non-luminal type, and even a 2-cm margin for skin surgical lines may result in positive margins. There are cases where CLALN after ALND should also be considered possible metastasis. In DLI recurrence, the skin excision margin line should be set very generously.


Interferon suppresses erythromycin metabolism in rats and human subjects.
Interferon down-regulates expression of cytochrome P-450 3A in male rats. This study explored the hypothesis that interferon therefore decreases the metabolism of drugs catalyzed by cytochrome P-450 3A. Initial experiments in male rats used microsomal erythromycin N-demethylase activity as a probe for cytochrome P-450 3A catalytic activity. After administration of rat interferon-gamma, erythromycin metabolism was impaired (53% of control; p < 0.01). This change correlated with the decline in cytochrome P-450 3A-dependent androstenedione 6 beta-hydroxylase activity, indicating that the decrease in erythromycin N-demethylase activity could be attributed to interferon-mediated suppression of cytochrome P-450 3A. We then used the [14C]N-methyl erythromycin breath test to assess the activity of hepatic cytochrome P-450 3A in rats and human subjects before and after a single dose of interferon. In rats, rat interferon-gamma decreased erythromycin metabolism to 57% of control (p < 0.005). In the human study, six patients with chronic active hepatitis C and four healthy controls were examined 20 to 26 hr after receiving a subcutaneous injection of human interferon-alpha 2b. Interferon produced a small decrease (median = 15%; range = 3% to 35%) in erythromycin metabolism (p < 0.05), as determined by 2-hr excretion of 14CO2 in the breath. Thus interferon-mediated suppression of cytochrome P-450 3A is less strong in human subjects than in male rats.(ABSTRACT TRUNCATED AT 250 WORDS)


The Pelagos Sanctuary for Mediterranean marine mammals: Marine Protected Area (MPA) or marine polluted area? The case study of the striped dolphin (Stenella coeruleoalba).
The concurrence of man-made pressures on cetaceans in the Mediterranean Sea is potentially affecting population stability and marine biodiversity. This needs to be proven for the only pelagic marine protected area in the Mediterranean Sea: the Pelagos Sanctuary for Mediterranean Marine Mammals. Here we applied a multidisciplinary tool, using diagnostic markers elaborated in a statistical model to rank toxicological stress in Mediterranean cetaceans. As a case study we analyzed persistent, bioaccumulative and toxic chemicals combined with a wide range of diagnostic markers of exposure to anthropogenic contaminants and genetic variation as marker of genetic erosion in striped dolphin (Stenella coeruleoalba) skin biopsies. Finally, a statistical model was applied to obtain a complete toxicological profile of the striped dolphin in the Pelagos Sanctuary and other Mediterranean areas (Ionian Sea and Strait of Gibraltar). Here we provide the first complete evidence of the toxicological stress in cetaceans living in Pelagos Sanctuary.


The thin electrolyte layer approach to corrosion testing of dental materials--characterization of the technique.
An innovative technique for corrosion testing of metallic dental materials is introduced. The thin electrolyte layer technique (TET) simulates the physical characteristics of the oral environment by employing a still, thin layer of an electrolyte, in contrast to bulk electrolyte techniques (BET) which utilize relatively large quantities of fluid. Limiting current density tests on a platinum electrode revealed a lower surface oxygen content for TET. Borate buffer (pH 6.8) was employed as an electrolyte. The effect of lower oxygen content in TET on passivation and polarization characteristics of 316L SS in 0.9% saline was investigated. The results revealed differences in the polarization resistance and open circuit potential development with time, as well as in anodic and cathodic polarization behavior. Lower O2 concentration in TET was attributed to different electrolyte convection characteristics under both testing conditions. Additionally, use of the TET resulted in better data reproducibility. Overall, this investigation led to a deeper understanding of the electrochemical processes inherent in thin electrolytes such as those found in the oral environment.


Protoporphyrin IX accumulation disrupts mitochondrial dynamics and function in ABCG2-deficient hepatocytes.
Targeted inhibition of multidrug ABCG2 transporter is believed to improve cancer therapeutics. However, the consequences of ABCG2 inhibition have not been systematically evaluated since ABCG2 is expressed in several organs including the liver. Here, we demonstrate that ABCG2-deficient hepatocytes have increased amounts of fragmental mitochondria accompanied by disruption of mitochondrial dynamics and functions. This disruption was due to ABCG2 knockout elevating intracellular protoporphyrin IX, which led to upregulation of DRP-1-mediated mitochondrial fission. The finding that ABCG2 deficiency can generate dysfunctional mitochondria in hepatocytes raises concerns regarding the systematic use of ABCG2 inhibitor in cancer patients.


A neuropeptide signal confers ethanol state dependency during olfactory learning in Caenorhabditis elegans.
Alcohol intoxication can impact learning and this may contribute to the development of problematic alcohol use. In alcohol (ethanol)-induced state-dependent learning (SDL), information learned while an animal is intoxicated is recalled more effectively when the subject is tested while similarly intoxicated than if tested while not intoxicated. When Caenorhabditis elegans undergoes olfactory learning (OL) while intoxicated, the learning becomes state dependent such that recall of OL is only apparent if the animals are tested while intoxicated. We found that two genes known to be required for signal integration, the secreted peptide HEN-1 and its receptor tyrosine kinase, SCD-2, are required for SDL. Expression of hen-1 in the ASER neuron and scd-2 in the AIA neurons was sufficient for their functions in SDL. Optogenetic activation of ASER in the absence of ethanol during learning could confer ethanol state dependency, indicating that ASER activation is sufficient to signal ethanol intoxication to the OL circuit. To our surprise, ASER activation during testing did not substitute for ethanol intoxication, demonstrating that the effects of ethanol on learning and recall rely on distinct signals. Additionally, intoxication-state information could be added to already established OL, but state-dependent OL did not lose state information when the intoxication signal was removed. Finally, dopamine is required for state-dependent OL, and we found that the activation of ASER cannot bypass this requirement. Our findings provide a window into the modulation of learning by ethanol and suggest that ethanol acts to modify learning using mechanisms distinct from those used during memory access.


Acute respiratory distress syndrome caused by salicylate intoxication.
Salicylate-induced acute respiratory syndrome (ARDS) is a well-known entity occurring in 35% of salicylate-intoxicated patient. Careful history taking, physical examination, arterial blood gas analysis, and measurement of serum salicylate concentration will lead to early recognition to initiate appropriate treatment.


Mapping of the linkage group GLO--Bf--HLA-B,C,A--PGM3. 1. Recombination frequencies.
This study confirms close linkage for the GLO--Bf--HLA-B,C,A complex, and proves linkage between the MHC loci and PGM3. For GLO--PGM3 and Bf--PGM3, respectively, loose linkage seems to be likely, and close linkage can be excluded. Our mapping data on chromosome 6 favor the hypothesis that the PGM3 locus is situated on the HLA-A side of the MHC complex. Yet fine-structure mapping should be confirmed only by segregation analyses in crossover families by testing simultaneously all of the relevant marker loci within uniform family material.


Sacral nerve stimulation enhances early intestinal mucosal repair following mucosal injury in a pig model.
Reducing intestinal epithelial barrier (IEB) dysfunctions is recognized as being of major therapeutic interest for various intestinal disorders. Sacral nerve stimulation (SNS) is known to reduce IEB permeability. Here, we report in a pig model that SNS enhances morphological and functional recovery of IEB following mucosal injury induced via 2,4,6-trinitrobenzenesulfonic acid. These effects are associated with an increased expression of tight junction proteins such as ZO-1 and FAK. These results establish that SNS enhances intestinal barrier repair in acute mucosal injury. They further set the scientific basis for future use of SNS as a complementary or alternative therapeutic option for the treatment of gut disorders with IEB dysfunctions such as inflammatory bowel diseases or irritable bowel syndrome. Intestinal epithelial barrier (IEB) dysfunctions, such as increased permeability or altered healing, are central to intestinal disorders. Sacral nerve stimulation (SNS) is known to reduce IEB permeability, but its ability to modulate IEB repair remains unknown. This study aimed to characterize the impact of SNS on mucosal repair following 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced lesions. Six pigs were stimulated by SNS 3 h prior to and 3 h after TNBS enema, while sham animals (n = 8) were not stimulated. The impact of SNS on mucosal changes was evaluated by combining in vivo imaging, histological and functional methods. Biochemical and transcriptomic approaches were used to analyse the IEB and mucosal inflammatory response. We observed that SNS enhanced the recovery from TNBS-induced increase in transcellular permeability. At 24 h, TNBS-induced alterations of mucosal morphology were significantly less in SNS compared with sham animals. SNS reduced TNBS-induced changes in ZO-1 expression and its epithelial pericellular distribution, and also increased pFAK/FAK expression compared with sham. Interestingly, SNS increased the mucosal density of neutrophils, which was correlated with an increase in trypsin and TGF-β1 levels compared with sham. Finally, SNS prevented the TNBS-induced increases in IL-1β and IL-4 over time that were observed with sham treatment. In conclusion, our results show that SNS enhances mucosal repair following injury. This study highlights novel mechanisms of action of SNS and identifies SNS as a new therapy for diseases with IEB repair disorders.


Silk fibroin promotes mineralization of gellan gum hydrogels.
Mineralization is a natural process leading to the formation of mineralized tissue such as bone. The chief mineral component of bone is hydroxyapatite (HAp), which is deposited using an organic template like fibrillar Collagen I under physiological condition. Fibrous silk fibroin is structurally homologous to collagen and acts as nucleation site for HAp mineralization when immersed in simulated body fluid (SBF) or fetal bovine serum (FBS), therefore, considered as popular bone regeneration biomaterial. Hence, the mineralization behavior of silk fibroin self-assembled gellan gum enriched 3D hydrogels is investigated under conditions closer to physiological ones using SBF as well as FBS, and also in presence of cells (e.g. human adipose tissue-derived stem cells, ASCs). Incorporation of silk fibroin induces the mineralization in acellular spongy-like hydrogels in composition dependent manner, confirmed by SEM-EDS analysis. In contrast, ASCs mediated mineralization is found in all hydrogel compositions of 3 weeks post-culture under osteogenic conditions as demonstrated by gene expression profile and Alizarin Red S staining. This is perhaps due to the co-existence of fibroin and FBS together induce cell-mediated mineralization. The blending of fibroin offers cheap alternative strategy to improve or guide the repair of mineralized tissue using gellan gum-based biomaterials.


Tools and Equipment for Managing Special Care Patients Anywhere.
This article describes many of the tools and equipment used by dental professionals to successfully treat special care patients in a variety of settings. Such equipment can be used in the dental office, operating room, hospital, surgical center, nursing home, private home, institution, hospice, and even in the field without electricity. Equipment discussed includes seating, laughing gas and sedation systems, body wraps and mouth props, lighting, radiographic exposure and imaging systems, dental isolation devices, and other tools the authors use.


SAGES white paper on the current state of community practice surgeons: re-discovering the value of our community practice surgeons.
The Community Practice (CP) surgeon is the first point of access to surgical care globally and performs the majority of procedures in the USA. CP surgeons include those of various practice models, locations and communities, education and training, and much more. It is a diverse group that drives quality, access to care, research, and innovation. The SAGES CP Committee was formed to better define the role and highlight the contribution of the CP surgeon, as well as advocate for the position of CP surgeons in our society. In 2018, a survey was distributed to the SAGES membership asking members to self-identify as either a Community Surgeon or Academic Surgeon. The majority (71%) of SAGES members surveyed self-identified as "Community Surgeons." This was in stark contrast to the distribution of Community versus Academic Surgeons in SAGES leadership (25% versus 75%, respectively). By better defining the characteristics and role of the CP, SAGES will be better informed on how to effectively engage with this large group within the society and increase its representation within the leadership. The CP Committee met on a biannual basis over a period of two years focusing on assessing their role in the SAGES organization. The committee members created the following initial goals: (1) define in a broad sense the characteristics of a CP Surgeon, (2) discuss and characterize the value of the CP surgeons, (3) highlight past and future areas of contributions of the group, and (4) delineate ways to engage and represent this subgroup. This manuscript is a culmination of the work of this committee while also serving as a way to support the initiatives and direction of SAGES leadership.


Role of phospholipids in the DDT-induced efflux of potassium in human erythrocytes.
Incubation of human erythrocytes for 1-2 h at 37 degrees C in a suspension of dipalmitoylphosphatidylcholine (DDPC) liposomes results in a phospholipid enrichment of erythrocyte membranes by 45-55% and a depletion of cholesterol by 19-24%. The enrichment by DPPC was time and concentration dependent. By contrast, dioleoylphosphatidylcholine (DOPC) liposomes were less effective in enriching the membranes with phospholipid and in depleting the membranes of cholesterol. Concomitantly, the DDT-induced efflux of K+ was reduced in the case of DPPC-enriched erythrocytes but enhanced in DOPC-enriched erythrocytes. These results suggest that DDT partitions more readily into the unsaturated than the saturated phospholipids of the erythrocyte membrane. It is concluded that the extent to which DDT affects the flux of K+ across the membrane is dependent on the fluidity of the lipid phase. We also report here a rapid method for cholesterol depletion of red blood cells in comparison to previously reported methods.


Clinic-based interventions to increase preexposure prophylaxis awareness and uptake among United States patients attending an obstetrics and gynecology clinic in Baltimore, Maryland.
Cisgender women account for 1 of every 5 new US HIV diagnoses, with most cases (85%) attributed to heterosexual contact. HIV preexposure prophylaxis is an effective prevention strategy; however, preexposure prophylaxis awareness and prescriptions among women are low. This study aimed to increase preexposure prophylaxis counseling and uptake among cisgender women attending obstetrics and gynecology clinics. The study included 3 obstetrics and gynecology clinics within a single health system in a high HIV prevalence region. There were 3 phases: baseline (the 3-month period before the clinical trial that included provider education and training of a registered nurse about preexposure prophylaxis), clinical trial (the 3-month period during which eligible patients were randomized to an active control or preexposure prophylaxis registered nurse intervention), and maintenance (the 3-month period after the trial ended). Electronic medical record clinical decision support tools were available to both arms during the clinical trial, which included best practice alerts, order sets, progress note templates, and written and video preexposure prophylaxis educational materials for patients. In the intervention arm, a preexposure prophylaxis nurse contacted and counseled patients and was equipped to prescribe preexposure prophylaxis. Moreover, this study evaluated the phases through the "reach, effectiveness, adoption, implementation, and maintenance" framework. The primary outcome of the study was effectiveness (eg, percentage of eligible patients with documented HIV prevention counseling in the electronic medical record or preexposure prophylaxis prescriptions). The secondary outcomes included reach (eg, percentage of best practice alerts that providers acted on or the percentage of eligible patients who spoke with the preexposure prophylaxis registered nurse), adoption (eg, percentage of eligible patients with a best practice alert that triggered or the percentage of eligible patients the preexposure prophylaxis registered nurse attempted to contact), and maintenance (eg, percentage of patients with documented HIV prevention counseling or preexposure prophylaxis prescriptions during the maintenance phase). There were 904 unique patients in all phases with a mean age of 28.8±7.7 years, and 416 patients (46%) were pregnant; moreover, 436 patients were randomized in the clinical trial phase. Concerning reach and adoption, best practice alerts were triggered for 100% of eligible encounters; however, the providers acted on 52% of them. The preexposure prophylaxis nurse attempted to contact every patient and successfully spoke with 81.2% of them in the preexposure prophylaxis registered nurse arm. Concerning effectiveness, there were significantly more patients counseled about preexposure prophylaxis in the preexposure prophylaxis registered nurse group than in the active control group (66.5% vs 12.3%, respectively; P<.001), although preexposure prophylaxis prescriptions were equivalent (P=1.0). Among the subgroup of patients who were counseled about preexposure prophylaxis, 18.5% of patients in the active control arm and 3.4% in the preexposure prophylaxis registered nurse arm were prescribed preexposure prophylaxis (P=.02). Concerning maintenance, clinical decision support tools alone resulted in preexposure prophylaxis counseling of 1.0% of patients during the maintenance phase vs 0.6% of patients during the baseline phase and 11.2% of patients during the clinical trial phase (P<.001). Preexposure prophylaxis prescriptions were not statistically different among the 3 phases (P=.096). A preexposure prophylaxis nurse effectively increased HIV prevention discussions but did not lead to more preexposure prophylaxis prescriptions than the preexposure prophylaxis-focused clinical decision support tools used by providers. The decrease in preexposure prophylaxis counseling after the trial phase suggests that persistent interventions are needed to maintain effects.


Radon remediation of a two-storey UK dwelling by active sub-slab depressurisation: effects and health implications of radon concentration distributions.
Radon concentration levels in a two-storey detached single-family dwelling in Northamptonshire, UK, were monitored continuously throughout a 5-week period during which active sub-slab depressurisation remediation measures were installed. Remediation of the property was accomplished successfully, with both the mean radon levels and the diurnal variability greatly reduced both upstairs and downstairs. Following remediation, upstairs and downstairs radon concentrations were 33% and 18% of their pre-remediation values respectively: the mean downstairs radon concentration was lower than that upstairs, with pre- and post-remediation values of the upstairs/downstairs concentration ratio, R(U/D), of 0.81 and 1.51 respectively. Cross-correlation between upstairs and downstairs radon concentration time-series indicates a time-lag of the order of 1 h or less, suggesting that diffusion of soil-derived radon from downstairs to upstairs either occurs within that time frame or forms a relatively insignificant contribution to the upstairs radon level. Cross-correlation between radon concentration time-series and the corresponding time-series for local atmospheric parameters demonstrated correlation between radon concentrations and internal/external pressure difference prior to remediation; this correlation disappears following remediation. Overall, these observations provide further evidence that radon concentration levels within a dwelling are not necessarily wholly determined by the effects of soil-gas advection, and further support the suggestion that, depending on the precise content of the building materials, upstairs radon levels, in particular, may be dominated by radon exhalation from the walls of the dwelling, especially in areas of low soil-gas radon.


EMD 52692 (bimakalim), a new potassium channel opener, attenuates luminol-enhanced chemiluminescence and superoxide anion radical formation by zymosan-activated polymorphonuclear leukocytes.
We investigated the relationship of potassium channel activation on modulation of oxidative respiratory bursts in canine neutrophils. Generation of superoxide anion radicals in opsonized zymosan-activated cells was determined using the technique of ferricytochrome c reduction. Preincubation of cells with the selective potassium channel opener, EMD 52692 (1-100 microM), attenuated superoxide anion radical production. Furthermore, EMD 52692 also produced a concentration-dependent inhibition of luminol-enhanced chemiluminescence by activated neutrophils. Glyburide, a selective antagonist of ATP-sensitive potassium channels, prevented the modulatory effect of EMD 52692 on both superoxide anion generation and luminol-enhanced chemiluminescence. The results suggest that ATP-sensitive potassium channels may play a significant role in regulating oxygen-derived free radical production in neutrophil-induced tissue injury.


Epigenetics and the plasticity of differentiation in normal and cancer stem cells.
Embryonic stem cells are characterized by their differentiation to all cell types during embryogenesis. In adult life, different tissues also have somatic stem cells, called adult stem cells, which in specific niches can undergo multipotent differentiation. The use of these adult stem cells has considerable therapeutic potential for the regeneration of damaged tissues. In both embryonic and adult stem cells, differentiation is controlled by epigenetic mechanisms, and the plasticity of differentiation in these cells is associated with transcription accessibility for genes expressed in different normal tissues. Abnormalities in genetic and/or epigenetic controls can lead to development of cancer, which is maintained by self-renewing cancer stem cells. Although the genetic abnormalities produce defects in growth and differentiation in cancer stem cells, these cells have not always lost the ability to undergo differentiation through epigenetic changes that by-pass the genomic abnormalities, thus creating the basis for differentiation therapy. Like normal stem cells, cancer stem cells can show plasticity for differentiation. This plasticity of cancer stem cells is also associated with transcription accessibility for genes that are normally expressed in different tissues, including tissues other than those from which the cancers originated. This broad transcription accessibility can also contribute to the behavior of cancer cells by overexpressing genes that promote cell viability, growth and metastasis.


In vitro bioaccessibility of vitamins B1, B2, and B3 from various vegetables.
B group vitamins, except folate, are involved in at least one step of cellular energy production. Vegetables are considered essential for a healthy diet plan. Vegetables significantly affect diet quality by contributing to the adequate intake of some B group vitamins. Our results demonstrated that the level of vitamins B1, B2, and B3 in the studied vegetables was in the range of 9-85 µg/100 g, 22-319 µg/100 g, and 459-3497 µg/100 g, respectively. However, it is fundamental to investigate the bioaccessibility of all vitamins to identify primary dietary sources. We observed that the average bioaccessibility values for vitamins B1 and B2 were 68.9% and 63.9%, respectively. The bioaccessibility of the nicotinic acid form of vitamin B3 was 40%, while the nicotinamide form was 33.9%. As revealed in this research, the bioaccessibilities of vitamins B1, B2, and B3 in vegetables were generally low in vitro.


Genomic parasites or symbionts? Modeling the effects of environmental pressure on transposition activity in asexual populations.
Transposable elements are DNA segments capable of persisting in host genomes by self-replication in spite of deleterious mutagenic effects. The theoretical dynamics of these elements within genomes has been studied extensively, and population genetic models predict that they can invade and maintain as a result of both intra-genomic and inter-individual selection in sexual species. In asexuals, the success of selfish DNA is more difficult to explain. However, most theoretical work assumes constant environment. Here, we analyze the impact of environmental change on the dynamics of transposition activity when horizontal DNA exchange is absent, based on a stochastic computational model of transposable element proliferation. We argue that repeated changes in the phenotypic optimum in a multidimensional fitness landscape may induce explosive bursts of transposition activity associated with faster adaptation. However, long-term maintenance of transposition activity is unlikely. This could contribute to the significant variation in the transposable element copy number among closely related species.


Isolation of High Purity Mouse Mesenchymal Stem Cells through Depleting Macrophages Using Liposomal Clodronate.
The use of mouse bone marrow mesenchymal stem cells (mBMSCs) represents a promising strategy for performing preclinical studies in the field of cell-based regenerative medicine; however, mBMSCs obtained via conventional isolation methods have two drawbacks, i.e., (i) they are heterogeneous due to frequent macrophage contamination, and (ii) they require long-term culturing for expansion. In the present study, we report a novel strategy to generate highly pure mBMSCs using liposomal clodronate. This approach is based on the properties of the two cell populations, i.e., BMSCs (to adhere to the plasticware in culture dishes) and macrophages (to phagocytose liposomes). Liposomal clodronate added during the first passage of whole bone marrow culture was selectively engulfed by macrophages in the heterogeneous cell population, resulting in their effective elimination without affecting the MSCs. This method allowed the generation of numerous high-purity Sca-1+CD44+F4/80- mBMSCs (> 95%) with just one passaging. Comparative studies with mBMSCs obtained using conventional methods revealed that the mBMSCs obtained in the present study had remarkably improved experimental utilities, as demonstrated by in vitro multilineage differentiation and in vivo ectopic bone formation assays. Our newly developed method, which enables the isolation of mBMSCs using simple and convenient protocol, will aid preclinical studies based on the use of MSCs.


Cardiometabolic risk markers during mood-stabilizing treatment: Correlation with drug-specific effects, depressive symptoms and treatment response.
Patients with bipolar disorder have higher rates of cardiometabolic comorbidities and mortality. Although guidelines emphasize the importance of cardiovascular monitoring, few studies characterized the cardiometabolic risk profile during treatment and their relation to symptomatology and treatment response. We analyzed data from two similar 24-weeks comparative effectiveness trials, with a combined sample of 770 participants randomized to two different lithium doses, quetiapine (300 mg/day), or standard treatment without lithium. Glucose, lipids and vital signs were measured before and after 24 weeks of treatment. We calculated several cardiovascular risk scores, assessed baseline correlations and compared the four treatment arms via multiple linear regression models. Higher cholesterol and LDL levels were associated with greater depression severity, showing differential correlations to specific symptoms, particularly agitation, low energy and suicidality. Those randomized to quetiapine showed a significant worsening of cardiometabolic markers during the 24-week trial. Neither baseline nor change in lipid levels correlated with differential treatment response. Study duration was short from the perspective of cardiometabolic risk markers, and all treatment arms included patients taking adjunct antipsychotics. The trials compared quetiapine to lithium, but not to other medications known to affect similar risk factors. Treatment with 300 mg/day quetiapine for 24 weeks, representing a short and common dose course, resulted in increased cardiometabolic risk markers, emphasizing the importance of monitoring during mood-stabilizing treatment. The symptom-specific associations are in line with previous studies in unipolar depression, suggesting a cardiometabolic-depression link that needs to be further studied in bipolar depression.


Comparison of efficacy of ivermectin and diethylcarbamazine against naturally infected Brugia malayi microfilaria in dogs.
Filarial parasites like Brugia pahangi and Brugia malayi can infect dogs. Adults of Brugia genus resides in the lymphatic system and microfilariae, in blood. There are increasing reports of detection of B. malayi microfilariae in dogs. A study was undertaken to compare the efficacy of repeated oral dosing of ivermectin (IVT) and diethylcarbamazine (DEC), individually and in combination against naturally infected B. malayi microfilariae in dogs. The species of the microfilariae was confirmed by acid phosphatase staining and polymerase chain reaction. The three treatment groups were 200 mcg/kg body weight IVT daily for 14 days (I), 6.6 mg/kg body weight DEC daily for 14 days (II) and IVT and DEC together in the same dose for a period of 5 days (III). Microfilarial status of the peripheral blood was assessed on the 0th, 7th, 14th and 21st day. Haematological parameters were measured on day zero and on the 21st day. Though, all the three treatment groups showed a reduction in the microfilarial concentration through the study period, complete absence of detectable microfilaremia was not noticed in any of the three groups by 21st day. Among the haematological parameters, statistically significant difference was observed in the post-treatment means of haemoglobin levels of group III when compared with group II. Since group III regime (IVT + DEC) was shorter and just as effective as the longer ones, it is considered superior to the other two.


Association of urinary N-domain Angiotensin I-converting enzyme with plasma inflammatory markers and endothelial function.
The aim of this study was to investigate the association between urinary 90 kDa N-domain Angiotensin I-converting enzyme (ACE) form with C-reactive protein (CRP) and homocysteine plasma levels (Hcy), urinary nitric oxide (NOu), and endothelial function (EF) in normotensive subjects. Forty healthy subjects were evaluated through brachial Doppler US to test the response to reactive hyperemia and a panel of blood tests to determine CRP and Hcy levels, NOu, and urinary ACE. They were divided into groups according to the presence (ACE90+) or absence (ACE90-) of the 90 kDa ACE, the presence (FH+) or absence (FH-) of family history of hypertension, and the presence or absence of these two variables FH+/ACE90+ and FH-/ACE90-. We found an impaired endothelial dilatation in subjects who presented the 90 kDa N-domain ACE as follows: 11.4% +/- 5.3% in ACE90+ compared with 17.6% +/- 7.1% in ACE90- group and 12.4% +/- 5.6% in FH+/ACE90+ compared with 17.7% +/- 6.2% in FH-/ACE90- group, P < 0.05. Hcy and CRP levels were statistically significantly lower in FH+/ACE90+ than in FH-/ACE90- group, as follows: 10.0 +/- 2.3 microM compared with 12.7 +/- 1.5 microM, and 1.3 +/- 1.8 mg/L compared with 3.6 +/- 2.0 mg/L, respectively. A correlation between flow-mediated dilatation (FMD) and CRP, Hcy, and NOu levels was not found. Our study suggests a reduction in the basal NO production confirmed by NOu analysis in subjects with the 90 kDa N-domain ACE isoform alone or associated with a family history of hypertension. Our data suggest that the presence of the 90 kDa N-domain ACE itself may have a negative impact on flow-mediated dilatation stimulated by reactive hyperemia.


Oat cell carcinoma of urinary bladder.
Oat cell carcinoma of the urinary bladder is extremely uncommon. There have been 52 reported cases in world literature to date. On light microscopic, histochemical, and ultrastructural grounds, these tumors are similar to oat cell carcinoma of the bronchus and other extrapulmonary oat cell carcinomas. Furthermore, they may be grouped together as a clinical entity characterized by an aggressive clinical course with early and extensive metastases, and partial remission with certain chemotherapeutic agents. We report a case of primary small cell carcinoma of the urinary bladder in a forty-two-year-old man and review previous reports with similar histology. The importance of establishing this diagnosis and the optimum forms of therapy are discussed.


Identification of Plasmodium knowlesi Merozoite Surface Protein-119 (PkMSP-119 ) novel binding peptides from a phage display library.
Plasmodium knowlesi, the fifth human malaria parasite, has caused mortality in humans. We aimed to identify P. knowlesi novel binding peptides through a random linear dodecapeptide phage display targeting the 19-kDa fragment of Merozoite Surface Protein-1 protein. rPkMSP-119 protein was heterologously expressed using Expresso® Solubility and Expression Screening System and competent E. cloni® 10G cells according to protocol. Three rounds of biopanning were performed on purified rPkMSP-119 to identify binding peptides towards rPkMSP-119 using Ph.D.™-12 random phage display library. Binding sites of the identified peptides to PkMSP-119 were in silico predicted using the CABS-dock web server. Four phage peptide variants that bound to PkMSP-119 were identified after three rounds of biopanning, namely Pkd1, Pkd2, Pkd3 and Pkd4. The sequences of both Pkd1 and Pkd2 consist of a large number of histidine residues. Pkd1 showed positive binding signal with 6.1× vs. BSA control. Docking results showed that Pkd1 and Pkd2 were ideal binding peptides for PkMSP-119 . We identified two novel binding peptides of PkMSP-119 , Pkd1 (HFPFHHHKLRAH) and Pkd2 (HPMHMLHKRQHG), through phage display. They provide a valuable starting point for the development of novel therapeutics.


Barriers and facilitators to physical activity participation in adults living with type 1 diabetes: a scoping review protocol.
The objective of this scoping review is to identify and map barriers to and facilitators of physical activity in adults living with type 1 diabetes. Physical activity is crucial to the day-to-day management of type 1 diabetes and in the prevention of diabetes-related complications. Despite these benefits, people living with type 1 diabetes have higher inactivity rates than those in the general population. Identifying barriers and facilitators to physical activity, specific to the type 1 diabetes population, may help explain this discrepancy. This scoping review will include articles describing adults aged 18 years or over, living with type 1 diabetes in any care setting. Included literature will focus on the key concepts under review: barriers to or facilitators of physical activity participation. Literature examining efficacy of strategies to manage blood glucose levels for physical activity will not be included. All sources of information will be reviewed, including peer-reviewed, published and unpublished literature. Database search limits will be applied to include articles written in English, involving human participants and published between 1996 and February 2019. Once all records are identified, duplicates will be removed. Remaining records will be subject to title and abstract screening where articles will be excluded if they clearly meet at least one exclusion criteria. All remaining full-text articles will be assessed for eligibility against inclusion and exclusion criteria. Included articles will undergo critical appraisal before being synthesized, charted and discussed.


High efficient key-insulated attribute based encryption scheme without bilinear pairing operations.
Attribute based encryption (ABE) has been widely applied for secure data protection in various data sharing systems. However, the efficiency of existing ABE schemes is not high enough since running encrypt and decrypt algorithms need frequent bilinear pairing operations, which may occupy too much computing resources on terminal devices. What's more, since different users may share the same attributes in the system, a single user's private key exposure will threaten the security and confidentiality of the whole system. Therefore, to further decrease the computation cost in attribute based cryptosystem as well as provide secure protection when key exposure happens, in this paper, we firstly propose a high efficient key-insulated ABE algorithm without pairings. The key-insulated mechanism guarantees both forward security and backward security when key exposure or user revocation happens. Besides, during the running of algorithms in our scheme, users and attribute authority needn't run any bilinear pairing operations, which will increase the efficiency to a large extent. The high efficiency and security analysis indicate that our scheme is more appropriate for secure protection in data sharing systems.


Unravelling the molecular and biochemical responses in cotton plants to biochar and biofertilizer amendments for Pb toxicity mitigation.
Over the past few years, there has been a rising interest in employing biochar (BC) and biofertilizers (BF) as a means of restoring soils that have been polluted by heavy metals. The primary objective of this study was to examine how the application of BC and BF affects the ability of cotton plants to withstand Pb toxicity at varying concentrations (0, 500, and 1000 mg/kg soil). The findings revealed that exposure to Pb stress, particularly at the 1000 mg/kg level, led to a decline in the growth and biomass of cotton plants. Pb toxicity triggered oxidative damage, impaired the photosynthetic apparatus, and diminished the levels of photosynthetic pigments. By increasing the expression of Rubisco-S, Rubisco-L, P5CR, and PRP5 genes and regulating proline metabolism, BC and BF increased the levels of proline and photosynthetic pigments and protected the photosynthetic apparatus. The application of BC and BF resulted in an upregulation of genes such as CuZnSOD, FeSOD, and APX1, as well as an increase in the activity of the glyoxalase system and antioxidant enzymes. These changes enhanced the antioxidant capacity of the plants and provided protection to membrane lipids from oxidative stress caused by Pb. The inclusion of BC and BF offered protection to photosynthesis and other essential intracellular processes in leaves by minimizing the transfer of Pb to leaves and promoting the accumulation of thiol compounds. This protective effect helped mitigate the negative impact of the toxic metal Pb on leaf function. By improving plant tolerance, reducing metal transfer, strengthening the antioxidant defense system, and enhancing the level of protective substances, these amendments show promise as valuable tools in tackling heavy metal pollution.


Cytotoxic effects of postharvest fungicides, ortho-phenylphenol, thiabendazole and imazalil, on isolated rat hepatocytes.
The cytotoxic effects of ortho-phenylphenol (OPP), imazalil (IMZ) and thiabendazole (TBZ) on isolated rat hepatocytes were investigated. Addition of IMZ and OPP to hepatocyte suspensions at a concentration of 0.75 mM resulted in acute cell death, accompanied by depletion of intracellular levels of glutathione and protein thiols. Both compounds rapidly depleted cellular ATP which consistently preceded the cell death. In addition, the cell death caused by IMZ was accompanied by the accumulation of intracellular malondialdehyde, indicating initiation of lipid peroxidation. During a 3-hr incubation period, TBZ did not affect these parameters. In mitochondria isolated from rat liver, IMZ and OPP impaired respiration related to oxidative phosphorylation. Based on these results, the order of toxic potency is IMZ > OPP > TBZ.


Epicardial Left Atrial Appendage Exclusion Reduces Blood Pressure in Patients With Atrial Fibrillation and Hypertension.
Percutaneous left atrial appendage exclusion (LAAE) has evolved as an alternative strategy for stroke prevention in atrial fibrillation (AF). Recent observational data have suggested that epicardial LAAE can have substantial impact on arrhythmia burden and hemodynamic profile. The authors aimed to study the impact of epicardial versus endocardial LAAE on systemic blood pressure in hypertensive AF patients. This was a prospective, nonrandomized study comparing 247 patients who underwent epicardial LAAE with 124 patients with endocardial exclusion. Clinical outcomes were measured at 3 months and 1 year. Primary outcome was improvement in systolic blood pressure (SBP) between both groups compared with baseline. Secondary outcome included changes in diastolic pressures (DBP), serum electrolytes, and creatinine. There was no significant difference in baseline SBP between epicardial and endocardial groups. SBP was significantly lower in the epicardial group both at 3 months (122 ± 11.8 mm Hg vs. 129.7 ± 8.2 mm Hg; p < 0.001) and 1 year (123 ± 11.6 mm Hg vs. 132.2 ± 8.8 mm Hg; p < 0.001) compared with the endocardial group. An adjusted multivariate linear mixed effects model demonstrated that epicardial LAAE significantly decreased SBP by 7.4 mm Hg at 3 months and by 8.9 mm Hg at 1 year (p < 0.0001). There was a trend toward lower DBP with epicardial LAAE at 3 months by 1.3 mm Hg (p = 0.2) and at 1 year by 1.8 mm Hg (p = 0.09). There was no significant difference in serum electrolytes and creatinine between both groups. In hypertensive AF patients, epicardial LAAE significantly decreases SBP both at 3 and 12 months compared with endocardial exclusion.


A case of congenital infantile fibrosarcoma of the right hand.
Congenital infantile fibrosarcoma is a rare soft-tissue malignant tumor that usually presents as a mass involving the arm or leg. We report a case of congenital fibrosarcoma on the right hand of a newborn infant. The patient was a 12-day-old male. From the time of delivery, he had a red tumor on his dorsal right hand. The tumor was initially associated with bleeding within a hemangioma and compressed. The course was observed. It appeared that the tumor might be malignant because it increased in size to 10 x 8 x 3 cm, and ulceration and bleeding appeared on the surface after 10 days. A biopsy suggested a fibrosarcoma. The tumor was resected 32 days after birth at the narrow margin in consideration of preservation of function. However, a recurrence was observed under the graft after 2 weeks and another resection was performed. Because the tumor had passed through the interosseous muscle and invaded the palm of the hand, the middle finger, ring finger, and little finger had to be amputated. One year and 7 months postoperatively, no recurrence or metastasis has been observed.


A Study of the Preparation and Properties of Antioxidative Copper Inks with High Electrical Conductivity.
Conductive ink using copper nanoparticles has attracted much attention in the printed electronics industry because of its low cost and high electrical conductivity. However, the problem of easy oxidation under heat and humidity conditions for copper material limits the wide applications. In this study, antioxidative copper inks were prepared by dispersing the nanoparticles in the solution, and then conductive copper films can be obtained after calcining the copper ink at 250 °C in nitrogen atmosphere for 30 min. A low sheet resistance of 47.6 mΩ/□ for the copper film was measured by using the four-point probe method. Importantly, we experimentally demonstrate that the electrical conductivity of copper films can be improved by increasing the calcination temperature. In addition, these highly conductive copper films can be placed in an atmospheric environment for more than 6 months without the oxidation phenomenon, which was verified by energy-dispersive X-ray spectroscopy (EDS). These observations strongly show that our conductive copper ink features high antioxidant properties and long-term stability and has a great potential for many printed electronics applications, such as flexible display systems, sensors, photovoltaic cells, and radio frequency identification.


Effect of obesity on spinal loads during load-reaching activities: A subject- and kinematics-specific musculoskeletal modeling approach.
Obesity has been associated to increase the risk of low back disorders. Previous musculoskeletal models simulating the effect of body weight on intervertebral joint loads have assumed identical body postures for obese and normal-weight individuals during a given physical activity. Our recent kinematic-measurement studies, however, indicate that obese individuals adapt different body postures (segmental orientations) than normal-weight ones when performing load-reaching activities. The present study, therefore, used a subject- and kinematics-specific musculoskeletal modeling approach to compare spinal loads of nine normal-weight and nine obese individuals each performing twelve static two-handed load-reaching activities at different hand heights, anterior distances, and asymmetry angles (total of 12 tasks × 18 subjects = 216 model simulations). Each model incorporated personalized muscle architectures, body mass distributions, and full-body kinematics for each subject and task. Results indicated that even when accounting for subject-specific body kinematics obese individuals experienced significantly larger (by ∼38% in average) L5-S1 compression (2305 ± 468 N versus 1674 ± 337 N) and shear (508 ± 111 N versus 705 ± 150 N) loads during all reaching activities (p < 0.05 for all hand positions). This average difference of ∼38% was similar to the results obtained from previous modeling investigations that neglected kinematics differences between the two weight groups. Moreover, there was no significant interaction effect between body weight and hand position on the spinal loads; indicating that the effect of body weight on L5-S1 loads was not dependent on the position of hands. Postural differences alone appear, hence, ineffective in compensating the greater spinal loads that obese people experience during reaching activities.


In Situ Soft X-ray Spectromicroscopy of Early Tricalcium Silicate Hydration.
The understanding and control of early hydration of tricalcium silicate (C₃S) is of great importance to cement science and concrete technology. However, traditional characterization methods are incapable of providing morphological and spectroscopic information about in situ hydration at the nanoscale. Using soft X-ray spectromicroscopy, we report the changes in morphology and molecular structure of C₃S at an early stage of hydration. In situ C₃S hydration in a wet cell, beginning with induction (~1 h) and acceleration (~4 h) periods of up to ~8 h, was studied and compared with ex situ measurements in the deceleration period after 15 h of curing. Analysis of the near-edge X-ray absorption fine structure showed that the Ca binding energy and energy splitting of C₃S changed rapidly in the early age of hydration and exhibited values similar to calcium silicate hydrate (C-S-H). The formation of C-S-H nanoseeds in the C₃S solution and the development of a fibrillar C-S-H morphology on the C₃S surface were visualized. Following this, silicate polymerization accompanied by C-S-H precipitation produced chemical shifts in the peaks of the main Si K edge and in multiple scattering. However, the silicate polymerization process did not significantly affect the Ca binding energy of C-S-H.


Clinical utility of contrast-enhanced three-dimensional ultrasound imaging with Sonazoid: findings on hepatocellular carcinoma lesions.
We investigated clinical utility of contrast-enhanced three-dimensional ultrasound (CE 3D US) imaging with contrast medium Sonazoid for demonstrating characteristic enhancement of hepatocellular carcinoma (HCC). Among 115 focal liver tumors undergoing CE 3D US imaging, 70 HCCs confirmed with contrast-enhanced multi-detector computed tomography, contrast-enhanced magnetic resonance imaging or histopathologic examination were retrospectively analyzed. CE 3D US imaging was performed using Autosweep 3D scan functionality in the early, middle and late phase, after bolus injection of 0.2 ml Sonazoid. The CE 3D tomographic images reconstructed in parallel slices and sonographic angiogram images were independently reviewed by two reviewers. Kappa values were used to assess inter-reviewers' agreement. TUI images showed most of HCCs were detected with intratumoral vessels and early tumor enhancement in the early phase, expressed homogenous or heterogeneous tumor enhancement in the middle phase, and became hypoechoic or isoechoic in the late phase. The kappa values in the early, middle and late phase for inter-reviewer agreements regarding the characteristic enhancement of tumors were 0.817, 0.774, and 0.785. In addition, TUI images demonstrated satellite foci and tortuous tumor vessels in three orthogonal planes. Sonographic angiogram reconstructed by different rendering modes showed the vessels and tumor stain in spatial view. The spatial configuration of anatomic structures was revealed on basis of both TUI and sonographic angiogram images. CE 3D US imaging, with spatial visualization, is clinically useful to exhibit the characteristic enhancement of HCC tumors objectively.


Inhibitory components from the buds of clove (Syzygium aromaticum) on melanin formation in B16 melanoma cells.
In the course to find a new whitening agent, we evaluated the methanol extract from bud of clove (Syzygium aromaticum) on melanin formation in B16 melanoma cells. Eugenol and eugenol acetate were isolated as the active compounds and showed melanin inhibition of 60% and 40% in B16 melanoma cell with less cytotoxicity at the concentration of 100 and 200 μg/mL, respectively. Furthermore, an essential oil prepared from the bud of clove, which contain eugenol and eugenol acetate as dominant components, showed melanin inhibition of 50% and 80% in B16 melanoma cells at the concentration of 100 and 200 μg/mL, respectively.


Five-year safety of tildrakizumab in patients with moderate-to-severe psoriasis from two phase 3 trials (reSURFACE 1 and reSURFACE 2): number needed to harm for occurrence of adverse events of special interest.
Five-year tildrakizumab safety data have been reported as exposure-adjusted incidence rates (EAIRs) of patients with events per 100 patient-years (PYs) of exposure. To present 5-year safety data from reSURFACE 1/2 phase 3 trials as EAIRs of events per 100 PYs of exposure, and the number needed to harm (NNH) for one adverse event of special interest (AESI) to occur. Pooled analysis from two randomized controlled trials in patients with moderate-to-severe plaque psoriasis (n = 1800). PSOLAR registry was used as safety reference data for NNH estimation. Rates of AESI with tildrakizumab were comparable with rates reported in PSOLAR. The NNH for one-year severe infection occurrence was 412 with tildrakizumab 200 mg, and negative for tildrakizumab 100 mg due to lower rates in reSURFACE trials; the NNH for malignancy was 990 for one year with tildrakizumab 100 mg (negative for tildrakizumab 200 mg); and the NNH for major adverse cardiovascular events was 355 for one year with tildrakizumab 200 mg (negative for tildrakizumab 100 mg). Tildrakizumab demonstrated a favorable safety profile over 5 years with low rates of AESI, comparable to those of the PSOLAR. Consequently, the NNH for AESI with tildrakizumab were very high or negative due to lower event rates for tildrakizumab.


Chemical composition of cardoon (Cynara cardunculus L. var. altilis) petioles as affected by plant growth stage.
In the present study petioles were collected from cultivated cardoon (Cynara cardunculus L. var altilis) plants in central Greece at sixteen harvest dates (samples P1-P16) and nine growth stages (principal growth stages (PGS) 1-9). The main objective of this study was to study the influence of petioles' growth cycle on their chemical composition. The lipidic content, fatty acids, free sugars, organic acids, and tocopherol composition were analyzed. A total of 27 fatty acids were identified, being the palmitic acid (C16:0, 12.42-50%) the most abundant, except for samples P4 and P5 where linoleic acid revealed the highest relative abundance (C18:2n6c, 46.5 and 39.05%, respectively). The α-, β-, and γ-tocopherols were the identified isoforms of vitamin E, with sample P1 recording the highest tocopherol content. The detected organic acids were oxalic, quinic, malic, citric, and fumaric acids with the highest content being observed in sample P1. Sucrose was the major sugar in all the analyzed samples, followed by fructose, glucose, trehalose, and raffinose. The obtained results allowed us to characterize the chemical composition of cardoon petioles throughout its growth cycle and, consequently, to valorize the species through the exploitation of underutilized plant tissues.


Isolation of a cDNA encoding a mouse TFIID subunit containing histone H4 homology.
A cDNA encoding a mouse transcription factor IID (TFIID) subunit, containing histone H4 homology, was cloned and sequenced. The predicted 678-amino-acid (aa) sequence of this molecule showed 97 and 41% identity to the human and Drosophila melanogaster homologues, respectively. Four putative direct repeats were found in the most highly conserved region in the central part of this protein.


RNAi targeting multiple cell adhesion molecules reduces immune cell recruitment and vascular inflammation after myocardial infarction.
Myocardial infarction (MI) leads to a systemic surge of vascular inflammation in mice and humans, resulting in secondary ischemic complications and high mortality. We show that, in ApoE(-/-) mice with coronary ligation, increased sympathetic tone up-regulates not only hematopoietic leukocyte production but also plaque endothelial expression of adhesion molecules. To counteract the resulting arterial leukocyte recruitment, we developed nanoparticle-based RNA interference (RNAi) that effectively silences five key adhesion molecules. Simultaneously encapsulating small interfering RNA (siRNA)-targeting intercellular cell adhesion molecules 1 and 2 (Icam1 and Icam2), vascular cell adhesion molecule 1 (Vcam1), and E- and P-selectins (Sele and Selp) into polymeric endothelial-avid nanoparticles reduced post-MI neutrophil and monocyte recruitment into atherosclerotic lesions and decreased matrix-degrading plaque protease activity. Five-gene combination RNAi also curtailed leukocyte recruitment to ischemic myocardium. Therefore, targeted multigene silencing may prevent complications after acute MI.


Argininosuccinic aciduria. A developmental and biochemical case study.
An infant with argininosuccinic aciduria was detected through the routine newborn screening program for inborn metabolic diseases and has been followed for over 7 years. Treatment consisting of restricted protein intake and arginine base supplementation was initiated at the age of 8 months and was continued intermittently. She maintained normal psychomotor development to the age of 3 years and currently at the age of 7.3 years, she has measured intelligence in the borderline range. She has had mild symptoms of cerebellar ataxia. Her physical growth has been below normal. Biochemical abnormalities are described. Special metabolic investigations and the effects of treatment are discussed.


Effect of an orthosis on the loads acting on a vertebral body replacement.
The spinal load reduction by an orthosis is still a matter of debate. Some studies predicted a load reduction while others found no effect. The aim of this study was to measure the in vivo effect of the Lumbo TriStep brace and the hyperextension orthosis medi 3C on the spinal implant loads. Telemeterized vertebral body replacements were implanted in 5 patients suffering from a severe fracture of the L1 or L3 vertebral body. The implant allows the measurement of 6 load components acting on it. For several activities during standing, sitting and walking, implant loads were measured in patients with and without an orthosis. The average resultant force on the vertebral body for 26 activities was reduced by 9% with the Lumbo TriStep brace, and by 19% with the hyperextension orthosis. The force reduction is usually more pronounced for activities performed during sitting than it is for those performed while standing. However, considerable inter- and intra-individual variation was observed. In several cases, the measured implant forces were even higher when the patients were wearing an orthosis. In some patients, for certain activities, an orthosis may reduce the force on a vertebral body replacement and thus on the anterior column of the spine. However, in other patients for the same activities, an orthosis may increase the force. The measurements do not allow a clear recommendation to wear an orthosis since the clinically relevant reduction of implant forces is unknown.


A generic RNA-pulsed dendritic cell vaccine strategy for renal cell carcinoma.
We present a generic dendritic cell (DC) vaccine strategy for patients with renal cell carcinoma (RCC) based on the use of RNA as a source of multiplex tumor-associated antigens (TAAs). Instead of preparing RNA from tumor tissue of each individual RCC patient, we propose to substitute RNA prepared from a well characterized highly immunogenic RCC cell line (RCC-26 tumor cells) as a generic source of TAAs for loading of DCs. We demonstrate here that efficient RNA transfer can be achieved using lipofection of immature DCs, which are subsequently matured with a cytokine cocktail to express high levels of MHC and costimulatory molecules as well as the chemokine receptor CCR7. Neither RNA itself nor the lipid component impacted on the phenotype or the cytokine secretion of mature DCs. Following RNA loading, DCs derived from HLA-A2-positive donors were able to activate effector-memory cytotoxic T lymphocytes (CTLs) specific for a TAA ligand expressed by the RCC-26 cell line. CTL responses to RNA-loaded DCs reached levels comparable to those stimulated directly by the RCC-26 tumor cells. Furthermore, DCs expressing tumor cell RNA primed naïve T cells, yielding T cell lines with cytotoxicity and cytokine secretion after contact with RCC tumor cells. RCC-26 cell lines are available as good manufacturing practice (GMP)-certified reagents enabling this source of RNA to be easily standardized and adapted for clinical testing. In addition, well defined immune monitoring tools, including the use of RNA expressing B cell lines, are available. Thus, this DC vaccine strategy can be directly compared with an ongoing gene therapy trial using genetically-engineered variants of the RCC-26 cell line as vaccines for RCC patients with metastatic disease.


Investigation of multiple factors which may contribute to vitamin D levels of bedridden pregnant women and their preterm neonates.
25-Hydroxyvitamin D (25-OH-D) is the marker, which indicates vitamin D levels. The aim of this study was to investigate the possible factors, which contribute to serum 25-OH-D levels in bedridden mothers and their preterm neonates. Twenty-six preterm neonates born during the period of 24-33 weeks of gestational age and 20 mothers (who experienced pregnancy complications) were recruited to the study. Five major results were obtained. (i) The 25-OH-D serum levels for preterm neonates and their mothers were found to possess strong correlation (ii) and both differed significantly in comparison with the optimal levels. (iii) An increase of mothers' 25-OH-D serum levels was associated with an increased possibility that the neonates would be measured to have normal 25-OH-D levels. (iv) Sex was not a key factor to neonates' 25-OH-D levels. (v) No correlation was found between mothers' 25-OH-D levels and their vitamin D3 supplement (400 IU/d during pregnancy). Due to insufficient exposure to sunlight and a diet not enriched with vitamin D, bedridden pregnant women suffer from vitamin D deficiency and pregnancy complications lead often to birth of preterm neonates with the same deficiency. Mothers should increase the total amount of vitamin D intake (food and supplement).


The REC1 gene of Ustilago maydis involved in the cellular response to DNA damage encodes an exonuclease.
Mutation in the REC1 gene of Ustilago maydis is known to lead to a complex phenotype with alterations in DNA repair, recombination, mutagenesis, meiosis, and cell division. The predicted product of the REC1 gene is a polypeptide of 522 amino acid residues with a molecular mass of 56,866 daltons, with no overall sequence homology to any other known protein. The open reading frame of the REC1 gene placed by itself in a U. maydis expression vector was found to be sufficient to complement the rec1 mutant. Overexpression of REC1 in Escherichia coli gave rise to the anticipated 57-kDa product together with a 3'-->5' exonuclease activity. This activity was only present in cells overexpressing REC1 and its characteristics were distinguishable from the major bacterial nucleases, but it had certain enzymatic features in common with epsilon, the proofreading exonuclease subunit of E. coli DNA polymerase III holoenzyme. To facilitate isolation of the protein product from bacteria, the REC1 gene was overexpressed from a vector that fused a hexa-histidine-leader sequence onto the amino terminus, enabling the isolation of the HisREC1 product on an immobilized metal ion affinity column. The His-REC1 protein co-eluted with the novel exonuclease activity. Alignment of the amino acid sequence of the REC1 gene product with the conserved proofreading exonuclease motifs of DNA polymerases indicated significant homology.


Limited compatibility between the RNA polymerase components of influenza virus type A and B.
Reassortants between type A and B influenza viruses have not been detected in nature, although both viruses co-circulate in human populations. One explanation for this may be functional incompatibility of RNA transcription and replication between type A and B viruses. To test this possibility, we constructed type A/B mosaic polymerase machinery, containing PB2, PB1, PA and nucleoprotein from each of the two virus types, and assessed their polymerase activities with a type A promoter in a reporter assay. Type B polymerase machinery containing homologous components was functional with the type A promoter albeit to various extents depending on the segments from which the regions downstream of the promoter sequence were derived, indicating functional compatibility between the type A promoter and B polymerase machinery. However, all of the A/B mosaic polymerase machinery, except that containing PA from a type A and the others from a type B virus strain, did not function with the type A promoter, indicating limited compatibility among polymerase components of both types. Taken together, these data suggest that incompatibility among components of the polymerase machinery for RNA transcription and replication alone is not responsible for the lack of heterotypic reassortants.


Persistence of Positive RT-PCR Results for Over 70 Days in Two Travelers with COVID-19.
The relation of continuing to test positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by reverse transcription real-time polymerase chain reaction (RT-PCR) to infectivity remains unclear, with numerous consequences. This report describes 2 patients with persistent viral detection by RT-PCR for 77 and 72 days, respectively, longer than other reported case-patients who were otherwise healthy.


Trabeculotome-guided deep sclerectomy. A pilot Study.
To describe and evaluate a new technique that helps identification and unroofing of Schlemm's canal during deep sclerectomy. A prospective, interventional case series. This pilot study was conducted on 15 eyes with various types of glaucoma. After dissecting the superficial scleral flap, the trabeculotome was inserted inside the Schlemm's canal. During deep flap dissection, a direct incision was made over the trabeculotomy to open and unroof Schlemm's canal. Five of the excised deep flaps were submitted for histologic examination. In 13 of the 15 eyes, the Schlemm's canal was properly identified and unroofed. Schlemm's canal endothelium was identified in all the examined specimens. The mean intraocular pressure was reduced from 26.66 +/- 4.54 mm Hg to 12.2 +/- 3.5 mm Hg at the end of a mean follow-up of 9.4 +/- 2.9 months. The insertion of the trabeculotome inside Schlemm's canal before dissection of the deep flap helped Schlemm's canal unroofing.


The potential of exosomes in immunotherapy of cancer.
Dendritic-cell-derived exosomes (DEX) secreted after dendritic cell loading with tumor peptides were found to mediate tumor rejection in mice. This observation prompted us to demonstrate that MHC class I/peptide complexes harbored onto exosomal membranes were capable of priming cytotoxic T cells and to mediate rejection of tumors expressing the relevant antigens. Moreover, DEX also promote NK cell activation in immunocompetent mice and NK cell-dependent antitumor effects. The first Phase I trial using DEX to immunize melanoma patients revealed the feasibility of DEX production in stage IV melanoma, their safety in long-term follow up and their bioactivity in vivo.


Paracrine exchanges of molecular signals between alginate-encapsulated pericytes and freely suspended endothelial cells within a 3D protein gel.
Paracrine signals, essential for the proper survival and functioning of tissues, may be mimicked by delivery of therapeutic proteins within engineered tissue constructs. Conventional delivery methods are of limited duration and are unresponsive to the local environment. We developed a system for sustained and regulated delivery of paracrine signals by encapsulating living cells of one type in alginate beads and co-suspending these cell-loaded particles along with unencapsulated cells of a second type within a 3D protein gel. This system was applied to vascular tissue engineering by placing human placental microvascular pericytes (PCs) in the particulate alginate phase and human umbilical vein endothelial cells (HUVECs) in the protein gel phase. Particle characteristics were optimized to keep the encapsulated PCs viable for at least two weeks. Encapsulated PCs were bioactive in vitro, secreting hepatocyte growth factor, an angiogenic protein, and responding to externally applied HUVEC-derived signals. Encapsulated PCs influenced HUVEC behavior in the surrounding gel by enhancing the formation of vessel-like structures when compared to empty alginate bead controls. In vivo, encapsulated PCs modulated the process of vascular self-assembly by HUVECs in 3D gels following implantation into immunodeficient mice. We conclude that alginate encapsulated cells can provide functional paracrine signals within engineered tissues.


Isolation, characterization and quantitation of chloral hydrate as a transient metabolite of trichloroethylene in man using electron capture gas chromatography and mass fragmentography.
A method for the quantitative determination of plasma concentrations of chloral hydrate by electron capture gas chromatography is described. Characterization by multiple ion monitoring confirms, for the first time, that chloral hydrate exists as a transient metabolite of trichloroethylene in man.


Heterogeneity in the low density lipoproteins of cholesterol fed African green monkeys (Cercopithecus aethiops).
The low density lipoproteins of cholesterol-fed African green monkeys were isolated by either preparative ultracentrifugation or agarose column chromatography, then radioiodinated and fractionated by agarose column chromatography. A marked variation was noted in the specific radioactivity of the column fractions regardless of the method used to obtain low density lipoproteins. The radioactivity in each fraction migrated with the beta-globluins, but when the fractions were mixed with carrier plasma, alpha-migrating radioactivity was also detected which was associated with the apo-A-I and apo-C of the high density lipoproteins. The uptake of these apoproteins by the high density lipoproteins reduced the variation in specific radioactivity among the low density lipoprotein fractions. By successive injections of the various 125I-labeled low density lipoprotein fractions into the same recipient, it was also possible to compare the kinetic behavior of the fractions. Large and small low density lipoprotein particles were metabolized differently, indicating that the low density lipoproteins from cholesterol-fed African green monkeys are heterogeneous.


Study on the mechanism of NLRP3 effect on the skeleton of de-ovalized mice.
Postmenopausal osteoporosis caused by estrogen deficiency affects millions of women worldwide. By influencing both osteoblast and osteoclast development, NOD-like receptor thermoprotein structural domain-associated protein 3 (NLRP3) is a key player in the etiology of osteoporosis (OP). The purpose of this research was to look into the mechanism of action of NLRP3 in osteoporosis caused by a lack of estrogen, highlighting that NLRP3 induces osteoblast pyroptosis and thus inflammatory responses in de-ovulated mice, thereby inhibiting osteogenic differentiation and participating in the development of osteoporosis. In de-ovulated mice, we found an enhanced inflammatory response and suppression of osteogenic activity. In vitro experiments, we found a significant increase in markers of cell pyroptosis and inflammatory responses and a significant decrease in markers of osteogenic differentiation in osteoblasts from de-ovulated mice. However, knockdown of the NLRP3 gene inhibited this cell pyroptosis and improved osteogenic differentiation of osteoblasts. Our findings indicate a potential therapeutic potential for the treatment of estrogen deficiency-induced osteoporosis by demonstrating the critical role that NLRP3 inflammatory vesicles and their downstream-mediated cellular pyroptosis play in bone differentiation.


[Treatment and patient information in the context of uncertain diagnosis].
An 82-year old female patient had an unclear gallbladder finding, and a laparoscopy was performed. The imaging techniques seemed to indicate gallbladder carcinoma infiltrating the liver. With the chosen method it was not possible to prove the actual existence of the cancer. The patient and her family were informed about the "benign" result and the patient was sent home. Three months later she died of gallbladder carcinoma, which was confirmed by liver aspiration during her last stay in hospital. The daughters were taken aback by the unexpected death of their mother. They declared that they would have spent the last three months differently, if they had been informed about the first suspected diagnosis and prognosis. Here we have a discussion about wasted chances. This article deals with the management of the fact that a diagnosed disease was ultimately fatal. It does not deal with the treatment of an advanced carcinoma of the gall-bladder with known lethal outcome. The approach can be twofold: a curative approach neglecting reality and patient communication on the one hand; a palliative approach with the goal of making the 'best' of the situation on the other. Regularly, after thorough clinical and technical examination we have evidence of cancer which we cannot actually prove by pathological confirmation. Very often these tumours are located in the pancreas and the biliary tract. When very old patients are concerned, decisions are even harder to take: how far are we to go with invasive examination methods and what will be the consequences after the existence of the cancer has been proven? What form should patient information take? In my opinion, all doctors are confronted with such situations almost daily.


Genetic Transformation of Fusobacterium nucleatum.
Fusobacterium nucleatum is a human periodontal pathogen that causes opportunistic infections. It has been implicated in preterm birth and has as a pathogen of colorectal cancer. However, it is a common member of the oral microbiota and can have a symbiotic relationship with its hosts. To date, studies of F. nucleatum have been hindered by a lack of effective genetic tools, and the transformation of F. nucleatum has not been investigated. In this chapter, protocols for the transformation of F. nucleatum strain 12230 using sonoporation are presented. We also include a genetic complementation protocol for a F. nucleatum knockout mutant.


Mechanism of action in a 4,5-diarylpyrrole series of selective cyclo-oxygenase-2 inhibitors.
Using semi-empirical AM1 calculation and 6.31G* basis sets, we have calculated the energy of the highest-occupied molecular orbital (E(HOMO)) for anti-inflammatory 4,5-diarylpyrroles which have been shown to have inhibitory activity on cyclooxygenase COX-2, an inducible enzyme expressed during inflammation. We have found a correlation between the E(HOMO) of a molecule and its COX-2 inhibition. However, no correlation was observed between E(HOMO) and the inhibition efficiency of cyclooxygenase-1 (COX-1), the constitutively expressed enzyme, protective to the organism. This result suggests that the inhibitions of the two isoforms follow different molecular mechanisms.


Correlations between Vancouver type of periprosthetic femur fracture and treatment outcomes.
Periprosthetic femur fractures are one of the most severe complications of total hip arthroplasty. The aim of this study was to assess outcomes of periprosthetic femur fractures based on the Vancouver classifications of fractures. Sixty-four patients were enrolled into the study. Type B1 fractures were the most common - 33 cases. In comparison to patients with type B3 fracture, patients with type B1 fracture significantly less often experienced postoperative pain. Study shown that in the context of both objective outcomes of the operation and patient satisfaction, the most unfavourable prognosis is after type B3.


Patient satisfaction with reduction mammaplasty: an early survey.
There is an abundant volume of literature available on the varying techniques of reduction mammaplasty, possible complications, and long-term evaluations from the surgeon's perspective. However, there is very little to be found on the patient's perspective and personal experience with this procedure during the perioperative and early postoperative period. This article deliberately concentrates on patients' experiences and impressions during the first and second postoperative months. A prospective study was undertaken to look at patients' personal experiences and satisfaction with different aspects of their reduction mammaplasty and their overall satisfaction with the procedure. Thirty-one consecutive patients were recruited during a six-month period and evaluated one month postoperatively. Twenty-seven of these patients were evaluated again two months postoperatively. The overall satisfaction rate was high: 80.6% for the one-month survey group and 88.9% for the two-month survey group. There was a high incidence of minor complications (38%) and a significant number of the patients (32.3% of the one-month survey group and 33.3% of the two-month survey group) felt the need for more preoperative discussion.


[Multi-mathematical modelings for compatibility optimization of Jiangzhi granules].
To investigate into the method of "multi activity index evaluation and combination optimized of mult-component" for Chinese herbal formulas. According to the scheme of uniform experimental design, efficacy experiment, multi index evaluation, least absolute shrinkage, selection operator (LASSO) modeling, evolutionary optimization algorithm, validation experiment, we optimized the combination of Jiangzhi granules based on the activity indexes of blood serum ALT, ALT, AST, TG, TC, HDL, LDL and TG level of liver tissues, ratio of liver tissue to body. Analytic hierarchy process (AHP) combining with criteria importance through intercriteria correlation (CRITIC) for multi activity index evaluation was more reasonable and objective, it reflected the information of activity index's order and objective sample data. LASSO algorithm modeling could accurately reflect the relationship between different combination of Jiangzhi granule and the activity comprehensive indexes. The optimized combination of Jiangzhi granule showed better values of the activity comprehensive indexed than the original formula after the validation experiment. AHP combining with CRITIC can be used for multi activity index evaluation and LASSO algorithm, it is suitable for combination optimized of Chinese herbal formulas.


RIG-I contributes to the innate immune response after cerebral ischemia.
Focal cerebral ischemia induces an inflammatory response that when exacerbated contributes to deleterious outcomes. The molecular basis regarding the regulation of the innate immune response after focal cerebral ischemia remains poorly understood. In this study we examined the expression of retinoic acid-inducible gene (RIG)-like receptor-I (RIG-I) and its involvement in regulating inflammation after ischemia in the brain of rats subjected to middle cerebral artery occlusion (MCAO). In addition, we studied the regulation of RIG-I after oxygen glucose deprivation (OGD) in astrocytes in culture. In this study we show that in the hippocampus of rats, RIG-I and IFN-α are elevated after MCAO. Consistent with these results was an increased in RIG-I and IFN-α after OGD in astrocytes in culture. These data are consistent with immunohistochemical analysis of hippocampal sections, indicating that in GFAP-positive cells there was an increase in RIG-I after MCAO. In addition, in this study we have identified n-propyl gallate as an inhibitor of IFN-α signaling in astrocytes. Our findings suggest a role for RIG-I in contributing to the innate immune response after focal cerebral ischemia.


Genes and susceptibility to leishmaniasis.
Leishmania are digenetic protozoa which inhabit two highly specific hosts, the sandfly where they grow as motile, flagellated promastigotes in the gut, and the mammalian macrophage where they grow intracellularly as non-flagellated amastigotes. Leishmaniasis is the outcome of an evolutionary 'arms race' between the host's immune system and the parasite's evasion mechanisms which ensure survival and transmission in the population. The spectrum of disease manifestations and severity reflects the interaction between the genome of the host and that of the parasite, and the pathology is caused by a combination of host and parasite molecules. This chapter examines the genetic basis of host susceptibility to disease in humans and animal models. It describes the genetic tools used to map and identify susceptibility genes, and the lessons learned from murine and human cutaneous leishmaniasis.


Unusual cold-induced disorders: cold-dependent dermatographism and systemic cold urticaria.
Two patients are described in whom new cold-induced disorders were defined. The patients presented with histories suggesting either dermatographism or typical cold urticaria; however, standard tests for each disorder failed to reproduce the patients' symptoms. One patient had dermatographism that was observable only upon chilling the skin subsequent to scratching it. The reaction was associated with detectable elevation of venous histamine levels coincident with the development of urticaria. The second patient had generalized urticaria that was induced by systemic rather than local cold challenge. The reaction was anaphylactoid in nature and was associated with systemic elevation of histamine levels. This patient was also mildly dermatographic, and skin reactivity was markedly augmented during an episode of generalized hives. These cold-dependent disorders should be included in the differential diagnosis of patients with histories suggestive of cold urticaria with or without a dermatographic component. Further evaluation is indicated when the standard tests for either of these disorders are negative.


[Malignant tumors of the salivary glands: early diagnosis, follow-up and therapy].
Problems of early recognition, postoperative care, and therapy were studied in 207 patients with malignant tumors of the salivary glands, who were treated from 1965 to 1975. Early recognition of these malignant tumors is hampered by the circumstance that only a very short time span is available for diagnosis of the highly malignant types and by the lack of knowledge about specific risk groups among the population. Important for postoperative care is the high rate of local tumor recurrence, especially of salivary-duct, mucoepidermoid, acinic-cell, and adenoid-cystic carcinomas, ranging from 71% to 83%. Late recurrence is frequent. There is also a close connection between local recurrence and lymphogenic or hematogenic formation of metastases. The therapy of choice for malignant tumors of the salivary glands is surgery and, depending on the tumor type, postoperative irradiation. Chemotherapy is at present still of minor importance. Histologic tumor type and tumor stage determine the extent of the surgical procedures, which range from lateral parotidectomy with preservation of the facial nerve to radical parotidectomy with or without reconstruction of the facial nerve. In spite of radical surgery, many malignomas of the salivary glands, particularly the adenoid-cystic carcinomas, tend to recur. In such cases the long-term prognosis is poor and cannot be improved by postoperative irradiation. Supplemented by the experience gained in postoperative care, we have summarized the guidelines along which these 207 salivary-gland malignomas were treated into a modified concept of therapy.


Microprocessor controlled trituration device for the dissociation of cardiac and other tissues.
The construction of an inexpensive microprocessor controlled piston/cylinder used in the dissociation of canine cardiac Purkinje fibres is described. The microprocessor, interfaced to a linear actuator, controls the motion of a piston using the higher level language "Tiny BASIC" and two machine language routines. Pressure changes generated by the piston are transferred to the lumen of a pipette. Movement of enzymatically-treated tissues through the pipette orifice facilitates dissociation into single cells. With this device, specific agitation protocols for different quantities and types of tissues are possible.


[Nonexponential diffusion extinction of spin echo from water protons in wheat root].
The method of NMR spin echo with the magnetic field pulse gradient was used for studying self-diffusion of water molecules in the radial root direction. It was shown on the basis of physiologopharmacological methods of modification of the object that the coefficients of water self-diffusion resulting from computer decomposition of nonexponential diffusion extinction of the relative echo amplitude in the root to components are related to the subcellular and supercellular organization (structure) of the root and reflect changes in water permeability of the two transport channels of plasmodesmas.


Origin of the arachidonic acid released post-mortem in rat forebrain.
To determine the origins of the arachidonic acid released post-mortem in brain tissue, [3H]arachidonic acid was injected by the intracerebro-ventricular route and radioactivity monitored in complex lipids and free arachidonic acid at various times after decapitation. The specific activity of the released arachidonic acid was close to that in the total phospholipid fraction and much lower than that of the neutral lipids. The phospholipid with the closest specific activity to the free arachidonic acid recovered at the end of the post-mortem period was phosphatidylinositol. Phosphatidylcholine showed a small but significant decrease in radioactivity post-mortem and could contribute 37% of the arachidonic acid released to the free fatty acid fraction. Arachidonic acid released in rat forebrain after decapitation thus comes from a mixture of phospholipids with phosphatidylinositol and phosphatidylcholine being the major source. Phosphatidylserine and phosphatidic acid did not make important contributions to the free arachidonic acid. In the microsomal fraction, the specific activity of the free arachidonic acid was very close to that in phosphatidylinositol.


A dynamic nuclear polarization strategy for multi-dimensional Earth's field NMR spectroscopy.
Dynamic nuclear polarization (DNP) is introduced as a powerful tool for polarization enhancement in multi-dimensional Earth's field NMR spectroscopy. Maximum polarization enhancements, relative to thermal equilibrium in the Earth's magnetic field, are calculated theoretically and compared to the more traditional prepolarization approach for NMR sensitivity enhancement at ultra-low fields. Signal enhancement factors on the order of 3000 are demonstrated experimentally using DNP with a nitroxide free radical, TEMPO, which contains an unpaired electron which is strongly coupled to a neighboring (14)N nucleus via the hyperfine interaction. A high-quality 2D (19)F-(1)H COSY spectrum acquired in the Earth's magnetic field with DNP enhancement is presented and compared to simulation.


Permeability of the rhesus monkey oocyte membrane to water and common cryoprotectants.
Successful cryopreservation of oocytes of the rhesus monkey (Macaca mulatta) would facilitate the use of this valuable animal model in research on reproduction and development, while providing a stepping stone towards human oocyte cryopreservation and the conservation of endangered primate species. To enable rational design of cryopreservation techniques for rhesus monkey oocytes, we have determined their osmotic and permeability characteristics in the presence of dimethylsulfoxide (DMSO), ethylene glycol (EG), and propylene glycol (PROH), three widely used cryoprotectants. Using nonlinear regression to fit a membrane transport model to measurements of dynamic cell volume changes, we estimated the hydraulic conductivity (L(p)) and cryoprotectant permeability (P(s)) of mature and immature oocytes at 23.5 degrees C. Mature oocyte membranes were most permeable to PROH (P(s) = 0.56 +/- 0.05 microm/sec) and least permeable to DMSO (P(s) = 0.24 +/- 0.02 microm/sec); the permeability to EG was 0.34 +/- 0.07 microm/sec. In the absence of penetrating cryoprotectants, mature oocytes had L(p) = 0.55 +/- 0.05 microm/min/atm, whereas the hydraulic conductivity increased to 1.01 +/- 0.10, 0.61 +/- 0.07, or 0.86 +/- 0.06 microm/min/atm when mature oocytes were exposed to DMSO, EG, or PROH, respectively. The osmotically inactive volume (V(b)) in mature oocytes was 19.7 +/- 2.4% of the isotonic cell volume. The only statistically significant difference between mature and immature oocytes was a larger hydraulic conductivity in immature oocytes that were exposed to DMSO. The biophysical parameters measured in this study were used to demonstrate the design of cryoprotectant loading and dilution protocols by computer-aided optimization.


[Evaluation of elderly patients with community-acquired pneumonia admitted to our hospital, who have neurologic disease, such as late effects of cerebral strokes in the past and medical history].
Hospitalized cases of community-acquired pneumonia (CAP) of the elderly with a history of neurologic diseases have not been examined in detail in the past. We extracted 15 cases with a history of neurologic disease (neurological history group, NH, all cases were over 70 years old), and 22 age-matched cases without a history of neurological diseases (non neurologic history group, non-NH) among 47 CAP patients who admitted to our hospital from home during the past year, and compared the two groups in terms of: (1) the score of severity of pneumonia by A-DROP system, (2) outcome (3) the duration of hospitalization, (4) the duration from end of treatment of antibiotic treatment to discharge, compared between the survival discharge cases out of two groups (11 vs 22 cases), (5) the bacteria that were detected. (1) In the NH group, the score was significantly higher than that in the non-NH group. (2) In the NH group, 4 cases died while none did in the non-NH group. (3) In the NH group, it was 30.8 +/- 22.8 (average +/- standard deviation) days, significantly higher than the 17.6 +/- 5.9 days in the non-NH group, (4) In the NH group, it was 16.6 +/- 14.1 days, which was significantly higher than the 6.7 +/- 4.8 days in the non-NH group. (5) In the NH group, there was a high rate of detection of Klebsiella pneumoniae, Escherichia coli and Serratia marcesscense. An original comprehensive plan of treatment and care support is necessary for NH because the NH is different from other CAP conditions.