Controlling release from encapsulated drug-loaded devices: insights from modeling the dissolution front propagation.
Dissolution of drug from its solid form to a dissolved form is an important consideration in the design and optimization of drug delivery devices, particularly owing to the abundance of emerging compounds that are extremely poorly soluble. When the solid dosage form is encapsulated, for example by the porous walls of an implant, the impact of the encapsulant drug transport properties is a further confounding issue. In such a case, dissolution and diffusion work in tandem to control the release of drug. However, the interplay between these two competing processes in the context of drug delivery is not as well understood as it is for other mass transfer problems, particularly for practical controlled-release considerations such as an encapsulant layer around the drug delivery device. To address this gap, this work presents a mathematical model that describes controlled release from a drug-loaded device surrounded by a passive porous layer. A solution for the drug concentration distribution is derived using the method of eigenfunction expansion. The model is able to track the dissolution front propagation, and predict the drug release curve during the dissolution process. The utility of the model is demonstrated through comparison against experimental data representing drug release from a cylindrical drug-loaded orthopedic fixation pin, where the model is shown to capture the data very well. Analysis presented here reveals how the various geometrical and physicochemical parameters influence drug dissolution and, ultimately, the drug release profile. It is found that the non-dimensional initial concentration plays a key role in determining whether the problem is diffusion-limited or dissolution-limited, whereas the nature of the problem is largely independent of other parameters including diffusion coefficient and encapsulant thickness. We expect the model will prove to be a useful tool for those designing encapsulated drug delivery devices, in terms of optimizing the design of the device to achieve a desired drug release profile.


Regulation of NF-κB by PML and PML-RARα.
Promyelocytic Leukemia (PML) is a nuclear protein that forms sub-nuclear structures termed nuclear bodies associated with transcriptionally active genomic regions. PML is a tumour suppressor and regulator of cell differentiation. We demonstrate that PML promotes TNFα-induced transcriptional responses by promoting NF-κB activity. TNFα-treated PML-/- cells show normal IκBα degradation and NF-κB nuclear translocation but significantly reduced NF-κB DNA binding and phosphorylation of NF-κB p65. We also demonstrate that the PML retinoic acid receptor-α (PML-RARα) oncofusion protein, which causes acute promyelocytic leukemia, inhibits TNFα induced gene expression and phosphorylation of NF-κB. This study establishes PML as an important regulator of NF-κB and demonstrates that PML-RARα dysregulates NF-κB.


10 years' neuroblastoma screening in Europe: preliminary results of a clinical and biological review from the Study Group for Evaluation of Neuroblastoma Screening in Europe (SENSE).
Between January 1986 and May 1996, 870,313 children were tested in European neuroblastoma (NB) screening programmes. Among these children, 82 cases of NB (age range 4-24 months, median 11 months) were detected by screening. 83% of the patients had localised NB and 17% were diagnosed with generalised NB (stage 4, 10%; stage 4s, 7%). Unfavourable biological markers (MYCN amplification, loss of heterozygosity (LOH) 1p36, DNA di/tetraploidy) were observed in 14% of 76 biologically examined cases. The median follow-up time of all the patients was 21.5 months (range 1-101 months). To date, 69 patients are in complete remission (CR) and 2 patients have died due to therapy (stage 4, 1 patient; stage 3, 1 patient with unfavourable markers). Apart from screened patients, 16 other patients with NB were found who had previously had a normal screening test, i.e. 'false negative' patients (age range 10-41 months, median 31.5 months). The median interval between screening and diagnosis was 24.5 months (range 6-35 months). 11 of the 'false negative' patients suffered from generalised NB (stage 4) and 5 had localised NB at diagnosis. Unfavourable biological markers were observed in 7/12 patients. 5 patients have died, 2 achieved partial remission and 9 CR. 9 of the 11 patients with unfavourable biological markers diagnosed due to NB screening are currently in CR. It is very likely that, among the patients without unfavourable biological markers, we detected tumours which may have regressed spontaneously. These children may have undergone 'unnecessary,' but unavoidable, diagnostic procedures and therapy. To reduce the number of 'false negative' patients, a later screening could be helpful and should be evaluated.


Reference field specification and preliminary beam selection strategy for accelerator-based GCR simulation.
The galactic cosmic ray (GCR) simulator at the NASA Space Radiation Laboratory (NSRL) is intended to deliver the broad spectrum of particles and energies encountered in deep space to biological targets in a controlled laboratory setting. In this work, certain aspects of simulating the GCR environment in the laboratory are discussed. Reference field specification and beam selection strategies at NSRL are the main focus, but the analysis presented herein may be modified for other facilities and possible biological considerations. First, comparisons are made between direct simulation of the external, free space GCR field and simulation of the induced tissue field behind shielding. It is found that upper energy constraints at NSRL limit the ability to simulate the external, free space field directly (i.e. shielding placed in the beam line in front of a biological target and exposed to a free space spectrum). Second, variation in the induced tissue field associated with shielding configuration and solar activity is addressed. It is found that the observed variation is likely within the uncertainty associated with representing any GCR reference field with discrete ion beams in the laboratory, given current facility constraints. A single reference field for deep space missions is subsequently identified. Third, a preliminary approach for selecting beams at NSRL to simulate the designated reference field is presented. This approach is not a final design for the GCR simulator, but rather a single step within a broader design strategy. It is shown that the beam selection methodology is tied directly to the reference environment, allows facility constraints to be incorporated, and may be adjusted to account for additional constraints imposed by biological or animal care considerations. The major biology questions are not addressed herein but are discussed in a companion paper published in the present issue of this journal. Drawbacks of the proposed methodology are discussed and weighed against alternative simulation strategies.


Silver-embedded screens in the intensive care unit. A new tool to control multi-drug resistant bacterial cross-transmission.
The purpose of this study was to assess the effectiveness of silver-embedded surfaces (BactiBlock®) to prevent surface colonization by multi-resistant bacteria (MRB) and to reduce the incidence of MRB colonization and infection in patients admitted to an intensive care unit (ICU). A 6-month prospective observational study in a 24-bed mixed ICU divided into two identical subunits (12 beds each) was designed. Seven solid mobile screens were placed in one of the subunits while in the other cloth screens remained. Solid screens were constructed with high-density polyethylene embedded in Bactiblock®. To evaluate the effectiveness of screens coated with Bactiblock®, number of MRB isolates on screens were compared for 6 months. Likewise, numbers of new patients and ICU-stays with MRB colonization in the two subunits were compared. One hundred forty screen samples were collected in 10-point prevalent days. MRB were detected on 28 (20.0%) samples. Over the 70 samples taken on cloth folding screens, MRB were detected in 25 (35.7%), while only 3 (4.3%) of the 70 samples taken on Bactiblock® screens were positive for MRB (p < 0.001). The unit with Bactiblock® screens presented fewer number of ICU stays with MRB colonization (27.8% vs 47.1%; p < 0.001). No significant differences were found in the global incidence of MRB nosocomial infection. The presence of Bactiblock® embedded in solid folding screens avoided MRB surface colonization and reduced MRB transmission to patients admitted to critical care units, proving to be an useful tool in the control of MRB.


Captopril and ramiprilat protect against free radical injury in isolated working rat hearts.
The protection of angiotensin converting enzyme (ACE) inhibitors, captopril and ramiprilat, against free radical-mediated myocardial injury were studied in isolated working rat hearts. Free radicals were generated by electrolysis of Krebs-Henseleit solution with 10 mA direct current for 1 min. Both captopril (360 mumol/l) and ramiprilat (12.5 mumol/l) significantly reduced the decrease of left ventricle dP/dt'max, coronary flow (CF), myocardial superoxide dismutase (SOD) and creatine kinase (CK) activities and the elevation of S-T segment of epicardial ECG as well as the rise of myocardial malondialdehyde (MDA) content caused by electrolysed perfusate. Captopril afforded a dose-dependent protection on cardiac functions with various concentrations of 45, 90, 180 and 360 mumol/l. Iloprost (30 nmol/l), a stable mimetic of prostacyclin, could also alleviate free radical-mediated myocardial injuries. All the beneficial effects of ramiprilat (12.5 mumol/l) were abolished by the administration of indomethacin (5 mumol). In contrast, captopril (90 mumol/l) still exhibited significant protective effects after indomethacin (9 mumol) was administered, though these protective effects were insignificantly weakened. In order to assess the role of sulfhydryl (-SH) group in the effects of captopril, a SH-containing drug S8 and a disulfide DG4, both are deficient in ACE inhibitory properties in vitro, were examined. Data showed that S8 (180 mumol/l) provided a significant protection while DG4 showed no protective effect. It is concluded that ACE inhibitors can protect against free radical-induced myocardial damage. Ramiprilat, a non-SH-containing ACE inhibitor, inhibits free radical-induced damages mainly by stimulation of prostacyclin synthesis and/or release. In addition to this effect, captopril, a SH-containing ACE inhibitor, may exert additional anti-free radical effects by a mechanism which is probably related to the sulfhydryl group.


Time series analysis as an emerging method for researching L2 affective variables.
In language studies, marked by a myriad of psychological and social, and linguistic factors, linear modeling fails to represent the creativity, irregularity and emergent patterns of behavior. To adequately represent the dynamicity and complexity of psychological or affective variables, time-sensitive non-linear modeling is needed, especially the time series analysis (TSA), which accommodates incompatibility over time. TSA is a mathematical framework that can effectively show whether and to what degree the measured time series represent nonlinear variation through time. TSA makes prediction or retrodiction of complex and dynamic phenomena possible in future or past and, thus, can significantly contribute to the unraveling of the nuanced changes in the progress of different learner-related constructs during learning a new language. The present paper, at first, offers an introductory overview of the TSA, and then pinpoints its technical features and procedures. Exemplary works of research in language studies will be reviewed next, followed by useful conclusive remarks about the subject. Finally, suggestions will be made for further investigation of language-related affective variables using this innovative method.


Acetylation of human TCF4 (TCF7L2) proteins attenuates inhibition by the HBP1 repressor and induces a conformational change in the TCF4::DNA complex.
The members of the TCF/LEF family of DNA-binding proteins are components of diverse gene regulatory networks. As nuclear effectors of Wnt/β-catenin signaling they act as assembly platforms for multimeric transcription complexes that either repress or activate gene expression. Previously, it was shown that several aspects of TCF/LEF protein function are regulated by post-translational modification. The association of TCF/LEF family members with acetyltransferases and deacetylases prompted us to investigate whether vertebrate TCF/LEF proteins are subject to acetylation. Through co-expression with p300 and CBP and subsequent analyses using mass spectrometry and immunodetection with anti-acetyl-lysine antibodies we show that TCF4 can be acetylated at lysine K₁₅₀ by CBP. K₁₅₀ acetylation is restricted to TCF4E splice variants and requires the simultaneous presence of β-catenin and the unique TCF4E C-terminus. To examine the functional consequences of K₁₅₀ acetylation we substituted K₁₅₀ with amino acids representing the non-acetylated and acetylated states. Reporter gene assays based on Wnt/β-catenin-responsive promoter regions did not indicate a general role of K₁₅₀ acetylation in transactivation by TCF4E. However, in the presence of CBP, non-acetylatable TCF4E with a K₁₅₀R substitution was more susceptible to inhibition by the HBP-1 repressor protein compared to wild-type TCF4E. Acetylation of K₁₅₀ using a bacterial expression system or amino acid substitutions at K₁₅₀ alter the electrophoretic properties of TCF4E::DNA complexes. This result suggests that K₁₅₀ acetylation leads to a conformational change that may also represent the mechanism whereby acetylated TCF4E acquires resistance against HBP1. In summary, TCF4 not only recruits acetyltransferases but is also a substrate for these enzymes. The fact that acetylation affects only a subset of TCF4 splice variants and is mediated preferentially by CBP suggests that the conditional acetylation of TCF4E is a novel regulatory mechanism that diversifies the transcriptional output of Wnt/β-catenin signaling in response to changing intracellular signaling milieus.


Impact of Secreted Protein Acidic and Rich in Cysteine (SPARC) Expression on Prognosis After Surgical Resection for Biliary Carcinoma.
Secreted protein acidic and rich in cysteine (SPARC) is a matricellular protein that influences chemotherapy effectiveness and prognosis. The aim of this study was to investigate whether SPARC expression correlates with the postoperative survival of patients treated with surgical resection for biliary carcinoma. SPARC expression in resected biliary carcinoma specimens was investigated immunohistochemically in 175 patients. The relationship between SPARC expression and prognosis after surgery was evaluated using univariate and multivariate analyses. High SPARC expression in peritumoral stroma was found in 61 (35%) patients. In all patients, stromal SPARC expression was significantly associated with overall survival (OS) (P = 0.006). Multivariate analysis revealed that high stromal SPARC expression was an independent risk factor for poor OS (HR 1.81, P = 0.006). Moreover, high stromal SPARC expression was independently associated with poor prognosis in a subset of 118 patients treated with gemcitabine-based adjuvant chemotherapy (HR 2.04, P = 0.010) but not in the 57 patients who did not receive adjuvant chemotherapy (P = 0.21). Stromal SPARC expression correlated with the prognosis of patients with resectable biliary carcinoma, and its significance was enhanced in patients treated with adjuvant gemcitabine-based chemotherapy.


Relevance of Erk1/2-PI3K/Akt signaling pathway in CEES-induced oxidative stress regulates inflammation and apoptosis in keratinocytes.
2-Chloroethyl ethyl sulfide (CEES) is a well-known chemical warfare agent that induces cellular stress in exposed individuals. However, molecular mechanisms of CEES-induced oxidative stress-mediated metabolic deregulation are not clearly elucidated. Here we investigated CEES-induced free radical production act as key functional mediators of metabolic stress via Erk1/2 mitogen-activated protein kinases (MAPKs) and phosphatidylinositol-3-kinase (PI3K/Akt) signaling cascades in keratinocytes. We observed that CEES exposure disrupts the cellular antioxidant defense capacities leading to increase in free oxygen and nitrogen radical accumulation in keratinocytes. These unusual cellular abnormalities initiate cellular stress via Erk1/2-PI3K/Akt signaling pathways. Biochemical tools were used to analyze the changes in metabolites including sulfur amino acids (SAAs), namely, L-glutathione (GSH) and L-cysteine (Cys), in the presence of selective inhibitors of reactive oxygen/nitrogen species (ROS/RNS), Erk1/2, or PI3K/Akt after CEES exposure. Importantly, these metabolite changes were accompanied by a decrease in the glycolytic flux, consistent with the observed decrease in 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2) concentration and these CEES-induced phenomena were attenuated by pretreatment of Erk1/2 or PI3-K/Akt inhibitors. On the other hand, CEES exposure disrupts the protein carbonylation (PC) and lipid peroxidation (LPO) in keratinocytes leading to inflammation, crash of the cell-cell communication, cell cycle deregulation, and apoptosis via Erk1/2-PI3K/Akt pathways. However, pretreatment of Erk1/2 or PI3K/Akt inhibitors attenuated the CEES action. Collectively, these results illustrated that accumulated free radicals act as key functional mediators for inflammation, and apoptosis via Erk1/2-PI3K/Akt regulatory signaling cascades induced by CEES exposure. Treatment of pharmacological Erk1/2-PI3K/Akt inhibitors attenuated the CEES-induced keratinocyte injury that may provide the basis for the development of therapeutic strategy to work against CEES exposure.


Atypical femur fractures in patients receiving bisphosphonate therapy: etiology and management.
Osteoporosis is a growing problem that is projected to affect more than 50% of American adults by 2020. Bisphosphonate therapy is currently the primary mode of treating osteoporosis in this population. While bisphosphonate therapy has been successful in increasing bone mineral density, data has shown an increased risk of atypical femur fractures with prolonged therapy. Atypical femur fractures are characterized by low-energy or atraumatic injuries that occur in the subtrochanteric region. They originate on the medial cortex, travel transversely, and typically have little or no comminution. Conservative therapy is indicated for patients with incomplete fractures without prodromal symptoms. Patients with incomplete fractures and significant prodromal symptoms or visible fracture line on radiographs, those who have failed conservative management, and those with complete fractures should be treated with intramedullary nail fixation. Evaluation should involve imaging of the contralateral femur. Teriparatide therapy may be considered for patients without contraindications. While the incidence of these fractures is low, it is likely that these rates will increase with the aging population and increased prevalence of patients being treated with bisphosphonate therapy.


Reactive oxygen species mediate ET-1-induced activation of ERK1/2 signaling in cultured feline esophageal smooth muscle cells.
Reactive oxygen species (ROS) have been shown to play a critical role in propagating the signals of several growth factors, peptide hormones, and cytokines, such as epidermal growth factor, insulin, and interleukin-1. We investigated a possible role for ROS generation in mediating the action of ET-1 on activation of ERK1/2 in cultured feline esophageal smooth muscle cells (ESMC). Confluent layers of ESMC were stimulated by 10nM ET-1; activation of ERK was examined by western blot analysis with phospho-specific antibodies of ERKs. ET-1 induced ERK1/2 phosphorylation in a dose- and time- dependent manner. ERK1/2 activation by ET-1 reached the maximal levels at 5min showing slight activation up to 20min, and then slowly declined. It was confirmed that the activation of ERK1/2 was reduced by MEK inhibitor PD98059. We observed the dose-dependent inhibitory effect of diphenyleneiodonium (DPI), an inhibitor of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase on the ET-1-enhanced ERK1/2 phosphorylation in ESMC. Pretreatment of ESMC with N-acetylcysteine, a ROS scavenger, also attenuated the ET-1-induced ERK1/2 activation. In addition, DPI significantly inhibited the ET-1- induced ROS production when ROS was measured as a function of DCF fluorescence. The results suggest that ROS might be critical mediators of the ET-1-induced ERK1/2 signaling events in ESMC.


[Disseminated nocardiosis in a child with acute lymphoblastic leukemia].
Nocardiosis is a rare infectious disease in children. We report here a disseminated nocardiosis in a child with acute lymphoblastic leukemia. The patient presented prolonged febrile neutropenia and nodular pneumopathy. Based on the amplification of a 16S rDNA, a PCR assay detected Nocardia sp. in the patient's bronchoalveolar lavage (BAL) fluid. Culture of BAL samples yielded Nocardia nova colonies after 2 weeks of incubation. Hepatic, splenic, renal and cerebral localisations were detected on extension checkup. trimethoprime-sulfamethoxazole and amikacine were started given the results of PCR assay, with a good response. Improvement of the patient's general condition led to complete chemotherapy under ciprofloxacine and ceftriaxone treatment, without nocardiosis reactivation. Nocardiosis is a rare complication in children with acute lymphoblastic leukemia. trimethoprime-sulfamethoxazole prophylaxis is widely used to prevent Pneumocystis jiroveci infection in children with haematologic malignancies. As Nocardia species are usually sensible, trimethoprime-sulfamethoxazole could play a role in Nocardia prophylaxis in such population. In our patient, compliance with trimethoprime-sulfamethoxazole had been low. Nocardia species are relatively fastidious growth bacteria and are difficult to isolate with classical bacteriological techniques. Molecular methods are now available, with a good sensitivity and fast results allowing to start an appropriate antibiotherapy before culture results, as early treatment is a major prognosis factor in nocardiosis. Nocardia infection should be suspected in case of nodular pneumopathy in immunocompromised children. An extension checkup should be performed to detect secondary localisations.


Chylomicron retention disease: A rare cause of chronic diarrhea.
Chylomicron retention disease (CRD) is a rare autosomal recessive hereditary hypocholesterolemic disorder. The disease most frequently presents in infants and is characterized by a lipid malabsorption syndrome with steatorrhea, chronic diarrhea, and growth retardation. The disease is characterized by normal fasting serum triglyceride levels combined with the absence of apolipoprotein (apo) B48 and chylomicrons after a fat load. In this report, we describe the clinical, laboratory, and histological data as well as the molecular DNA analysis of a 12-month-old girl from Tunisia with CRD. The patient was treated with a low-fat diet and fat-soluble vitamin supplementation resulting in significant improvement.


Differential inhibition of two proteolytic activities in bovine lens neutral-proteinase preparations.
Hydrolysis of carbobenzoxy-Leu-Leu-Glu 2-naphthylamide by bovine lens neutral-proteinase preparations is not affected by the esterase inhibitor di-isopropyl fluorophosphate, whereas hydrolysis of carbobenzoxy-Gly-Gly-Leu p-nitroanilide is completely inhibited. Hydrolysis of alpha-crystallin, a lens structural protein, can be inhibited by only 50% after prolonged treatment with di-isopropyl fluorophosphate. These data suggest that the lens neutral-proteinase preparation contains at least two enzymes, one of which may be a serine proteinase. This may account, in part, for the previously observed complex response of the preparation to inhibitors.


Current development of 1,2,3-triazole derived potential antimalarial scaffolds: Structure- activity relationship (SAR) and bioactive compounds.
Malaria is among one of the most devastating and deadliest parasitic disease in the world claiming millions of lives every year around the globe. It is a mosquito-borne infectious disease caused by various species of the parasitic protozoan of the genus Plasmodium. The indiscriminate exploitation of the clinically used antimalarial drugs led to the development of various drug-resistant and multidrug-resistant strains of plasmodium which severely reduces the therapeutic effectiveness of most frontline medicines. Therefore, there is urgent need to develop novel structural classes of antimalarial agents acting with unique mechanism of action(s). In this context, design and development of hybrid molecules containing pharmacophoric features of different lead molecules in a single entity represents a unique strategy for the development of next-generation antimalarial drugs. Research efforts by the scientific community over the past few years has led to the identification and development of several heterocyclic small molecules as antimalarial agents with high potency, less toxicity and desired efficacy. Triazole derivatives have become indispensable units in the medicinal chemistry due to their diverse spectrum of biological profiles and many triazole based hybrids and conjugates have demonstrated potential in vitro and in vivo antimalarial activities. The manuscript compiled recent developments in the medicinal chemistry of triazole based small heterocyclic molecules as antimalarial agents and discusses various reported biologically active compounds to lay the groundwork for the rationale design and discovery of triazole based antimalarial compounds. The article emphasised on biological activities, structure activity relationships, and molecular docking studies of various triazole based hybrids with heterocycles such as quinoline, artemisinins, naphthyl, naphthoquinone, etc. as potential antimalarial agents which could act on the dual stage and multi stage of the parasitic life cycle.


Clinical Implementation of Total-Body PET/CT at University of California, Davis.
This work outlines the clinical implementation of total-body PET/CT from the time of installation at UC Davis EXPLORER Molecular Imaging Center in May 2019 to date (September 2020). This process has not been without challenges: logistical, technical, and medical problems were all encountered and are discussed. The mutual learning/teaching engagement among nuclear medicine technologists, research staff, radiologists, and referring physicians has been the key to the past year's achievements; these include development of ground-breaking clinical protocols for a range of Food and Drug Administration-approved radiotracers, and the attainment of a measure of expertise in clinical total-body PET/CT.


50 Hz magnetic fields of varying flux intensity affect cell shape changes in invertebrate immunocytes: the role of potassium ion channels.
The effect induced by exposure to 50 Hz magnetic fields (MFs) in immunocytes from the mussel Mytilus galloprovincialis is evaluated. The whole animal was exposed for 15 and 30 min to MF intensities ranging from 200 to 1,000 microT. The changes in the cellular shape of immunocytes, expressed as shape factor (SF), were studied at different times after addition of the chemotacting substance N-formyl-Meth-Leu-Phe (fMLP). Results show that MFs provoke differing delays in fMLP-induced cellular shape changes: 200 microT are ineffective, while levels from 300 microT upwards cause a significant increase in immunocyte SF values compared to controls. Reactivation of the cells is possible up to an intensity of 600 microT. The use of PCO 400, an opener of ATP-sensitive K+ channels, shows that potassium channels are involved in the effect of MFs on M. galloprovincialis immunocytes.


Localization of vasoactive intestinal peptide- and peptide histidine isoleucine amide-like immunoreactivities in the rat superior cervical ganglion and its nerve trunks.
Electrical stimulation of the preganglionic cervical sympathetic trunk causes an increase in dopa synthesis in the postganglionic neurons in the superior cervical ganglion (SCG). This transsynaptic biochemical effect can be blocked only partially by cholinergic antagonists, suggesting the involvement of a noncholinergic preganglionic sympathetic neurotransmitter(s). A survey of a large number of possible candidates for this neurotransmitter revealed that, in addition to cholinergic agonists, only a small group of peptides (all members of the secretin-glucagon family) stimulated dopa synthesis in the SCG. The effective peptides included vasoactive intestinal peptide (VIP), peptide histidine isoleucine amide (PHI), and secretin. Consequently we looked for the presence of immunoreactivities for these three peptides in the SCG. VIP- and PHI-like immunoreactive fibers were found in the SCG and in its major pre- and postganglionic nerve trunks. The distributions of the two immunoreactivities were very similar. Immunoreactive fibers were seen both singly and in bundles. In some instances, fibers were found apposed to neuronal cell bodies in the ganglion, and occasionally dense plexuses of fibers were found surrounding the neurons. In addition, punctate immunoreactive profiles were found apposed to the neurons in what appeared to be terminal fields. A small number of immunoreactive neuronal cell bodies were also seen in the ganglion. In a few instances, it was possible to establish, in serial sections, that the same cell body was immunostained with both VIP and PHI antisera. No secretin like-immunoreactive fibers or cells were observed. The presence of VIP-like and PHI-like-immunoreactive fibers in the cervical sympathetic trunk and in the SCG strengthens the possibility that these peptides, or a related molecule(s), serve as preganglionic neurotransmitters in this ganglion.


Calcium environment in encrusting deposits from urinary catheters investigated by interpretation of EXAFS spectra.
Extended x-ray absorption fine structure (EXAFS) spectra were recorded, above the K absorption edge of Ca, from 10 encrusted urinary (Foley) catheters obtained from 10 different patients. The presence of poorly crystalline apatite was demonstrated in all the deposits, even though this phase could not be detected in the x-ray powder diffraction patterns of most samples because of the obscuring effect of struvite, which was also present. Furthermore, the EXAFS spectra could be interpreted to yield quantitative information on the average short-range structure surrounding the Ca2+ ions in the poorly crystalline apatite.


Photon transmission tomography techniques for screening organs in biological specimen banking.
Biological specimen banking methodology dictates that when a number of specimens of a particular type are available then the selected specimen be representative of the population in question. It is apparent though that this sorting process may present a formidable task which can be facilitated by employing tomographic transmission techniques using photons. Thus it is possible to discriminate between and discern abnormalities in specimens by obtaining the distribution of the photon linear attenuation coefficient, a function of atomic number and physical density, in sections through the specimen whilst maintaining its integrity. Fresh and freeze-dried specimens of porcine kidney were initially examined under differing temperature conditions using a single point photon transmission method, which was not successful due to the heterogeneity of the tissues. Whereas photon transmission tomography is shown to be well suited in testing for homogeneity, highlights the importance of scanning before cryogenic treatment and provides the possibility of monitoring tissue status over storage period by observing small changes in linear attenuation coefficient.


Asynchronous development of electrical remodeling and cardiac hypertrophy in the complete AV block dog.
Left ventricular hypertrophy has been associated with the prolongation of QT-time, and an increased risk of ventricular arrhythmias. The renin angiotensin system has been implicated in the development of ventricular hypertrophy. At 5 weeks complete AV block (CAVB) in the dog, there is: (1) biventricular hypertrophy associated with a transient activation of components of the renin angiotensin system, (2) increased APD, more pronounced in the left than in the right ventricle leading to spatial dispersion of repolarization, and (3) enhanced susceptibility to drug-induced torsade de pointes arrhythmias. To investigate whether these remodeling processes develop in parallel, time dependency was assessed in absence or presence of the AT1 receptor-blocker Irbesartan. Dogs in sinus rhythm, 2 and 5 weeks CAVB were compared to dogs chronically treated with Irbesartan (30 mg/kg BID). Endocardial monophasic APD of left and right ventricle was measured and susceptibility to torsade de pointes was tested by infusing Dofetilide (0.025 mg/kg/5'). Hypertrophy was determined by relating heart-to-body weight at sacrifice. Left ventricular APD had increased more than right ventricular APD at 2 and 5 weeks CAVB, leading to an increase in spatial dispersion. At that time torsade de pointes were evocable in the majority of the dogs. Hypertrophy had only developed completely at 5 weeks CAVB. Irbesartan had no effect on electrical and structural parameters or on arrhythmogenicity. In the CAVB dog ventricular hypertrophy is not a prerequisite for electrical remodeling or drug-induced torsade de pointes, and the AT1-receptor has no dominant role in the completion of these remodeling processes.


Post-traumatic rotational deformity of the lower leg.
After a small series of patients with post-traumatic rotational deformity of the tibia treated by corrective osteotomy, we describe some diagnostic features and our indications for correction. Osteotomy must be considered for external rotation deformity of 20 degrees or more, and in cases of internal rotation deformity of over 15 degrees.


Protocol for measuring NLRC4 inflammasome activation and pyroptosis in murine bone-marrow-derived macrophages.
NLR family CARD domain containing protein 4 (NLRC4) inflammasome activation and the associated pyroptosis are critical for protection against infection by bacterial pathogens. This protocol presents a detailed procedure to activate and measure NLRC4 inflammasome activation and pyroptosis upon Salmonella Typhimurium infection. The techniques can be adapted to monitoring the activation of other types of inflammasomes and pathogenic stimuli. For comprehensive details on the use and execution of this protocol, please refer to Dong et al. (2021).


Single-cell RNA landscape of cell heterogeneity and immune microenvironment in ligation-induced vascular remodeling in rat.
Vascular remodeling is a common pathological basis for cardiovascular diseases. Although both immune and non-immune cells have been suggested to contribute to this process, the complex cellular heterogeneity and intercellular interactions remain largely uncharacterized. In this study, we simulated early and late vascular remodeling by ligating the rat carotid artery for 1 week and 4 weeks, respectively. Using single-cell RNA-sequencing, we characterized gene expression signatures and driver signals of major cell types involved in vascular remodeling. Focused analysis revealed a novel sub-population of Selenbp1hi smooth muscle cells (SMCs) associated with vascular remodeling. Results of intercellular communication analyses predicted several ligand-receptor pairs between immune cells with SMCs and endothelial cells (ECs), implicating SMCs apoptosis and repair, ECs aging and inflammatory responses. We present a comprehensive single-cell atlas of vascular cells in early and late stages of ligated rat carotid artery, providing valuable insights into the understanding of the initiation and progression of vascular remodeling.


Assessment of presumed small-molecule ligands of telomeric i-DNA by biolayer interferometry (BLI).
Biolayer interferometry (BLI) and circular dichroism (CD) spectroscopy were used to investigate the interaction between previously reported i-motif DNA (i-DNA) ligands and folded or unfolded i-DNA in acidic (pH 5.5) and near-neutral (pH 6.5) conditions. We observed that although several ligands, in particular macrocyclic bis-acridine (BisA) and pyridostatin (PDS), showed good affinities for the telomeric i-motif forming sequence, none of the ligands displayed selective interactions with the i-DNA structure nor was able to promote its formation.


An unusually stable trinuclear manganese(II) complex bearing bulk carboxylic radical ligands.
A trinuclear MnII cluster based on the versatile polychlorinated triphenylmethyl carboxylic radical ligand has been characterized by X-ray crystallography and magnetic measurements, representing a rare example of a MnII trinuclear linear structure with six open-shell ligands.


Burns management and junior staff--what do they know?
This study examines the ability of junior doctors to initiate the management of burned patients. One hundred and twenty-four junior doctors were assessed using a questionnaire. Eighty per cent of the sample had had undergraduate lectures on the subject and 43 per cent had experience of managing patients with major burns. Despite this only 3 per cent could correctly carry out all the steps necessary to estimate the fluid requirements of a burned patient. Theoretical knowledge of the 'Rule of Nines' was adequate but 10 per cent of the sample made mathematical errors when supplied with a burns formula and the appropriate values. We suggest that postgraduate instruction be given to junior staff and that burns charts include details of a burns formula and an illustrative example of the calculation required.


Tourism dependence and poverty alleviation thresholds in Chinese ethnic tourism.
Questions persist on the relationship between tourism dependence and economic growth in ethnic tourism areas. This study addresses such gaps by constructing a threshold regression model based on socio-economic data from 2006 to 2019 for nine sites in Enshi Prefecture of central China. ArcGIS and other open-source data were also used to visualize changing tourism resources in the region. Findings suggest that tourism dependence (the ratio of tourism-based GDP to overall GDP) significantly promotes economic growth in ethnic minority areas. However, the positive influence of tourism dependence on economic growth appears dynamic and non-linear - rising at first before falling when tourism dependence exceeded a threshold of 34%, with effects varying by site and year. Methods and findings make crucial theoretical contributions to understanding tourism dependence and poverty alleviation linkages. This paper also highlights the importance of political support and balanced investment in diverse industries to minimize decreasing returns beyond tourism dependence thresholds in destinations worldwide.


Magnetic parameters as proxies for anthropogenic pollution in water reservoir sediments from Mexico: An interdisciplinary approach.
We assess the element pollution level of water reservoir sediments using environmental magnetism techniques as a novel approach. Although "La Purísima" Water Reservoir is an important source for multiple activities (e.g. recreational, fishing and agricultural) in Guanajuato state, it has been receiving for the last centuries a high load of pollutants by mining extraction, urbanization and land-use change from the Guanajuato Hydrological Basin. The analyses of environmental magnetism, geochemistry, X-ray energy dispersive spectroscopy, scanning electron microscopy and multivariate methods were applied to study sediments from the reservoir and basin. Accordingly, they indicate the presence of iron oxides (magnetite and hematite) and iron sulfides (pyrite and greigite), which evidences relevant differences in particle size and concentration within the water reservoir (median mass-specific magnetic susceptibility χ = 23.2 × 10-8 m3/kg), as well as with respect to the river basin sediments (median χ = 88.8 × 10-8 m3/kg). The highest enrichment factor EF values (median values of EF = 2-10 for As, Co, Ba, Cu, Cd, Ni and EF > 20 for S) are mainly associated with historical mining activities that have led to an enrichment of potentially toxic elements on these water reservoir sediments. We propose the use of concentration and grain size dependent magnetic parameters, i.e. χ, remanent magnetizations and anhysteretic ratios ARM/SIRM and χARM/χ, as proxies for Ba, Co, Cr, Ni, P and Pb pollution in these river and water reservoir sediments. Such parameters allow to evaluate this sedimentary environment, and similar ones, through useful and convenient proxies.


Molar optimization versus delayed reinforcement as explanations of choice between fixed-ratio and progressive-ratio schedules.
In a discrete-trials procedure, pigeons chose between a fixed-ratio 81 schedule and a progressive-ratio schedule by making a single peck at the key correlated with one or the other of these schedules. The response requirement on the progressive-ratio schedule began at 1 and increased by 10 each time the progressive-ratio schedule was chosen. Each time the fixed-ratio schedule was chosen, the requirement on the progressive-ratio schedule was reset to 1 response. In conditions where there was no intertrial interval, subjects chose the progressive-ratio schedule for an average of about five consecutive trials (during which the response requirement increased to 41), and then chose the fixed-ratio schedule. This ratio was larger than that predicted by an optimality analysis that assumes that subjects respond in a pattern that minimizes the response-reinforcer ratio or one that assumes that subjects respond in a pattern that maximizes the overall rate of reinforcement. In conditions with a 25-s or 50-s intertrial interval, subjects chose the progressive-ratio schedule for an average of about eight consecutive trials before choosing the fixed-ratio schedule. This change in performance with the addition of an intertrial interval was also not predicted by an optimality analysis. On the other hand, the results were consistent with the theory that choice is determined by the delays to the reinforcers delivered on the present trial and on subsequent trials.


Effects of conjugated linoleic acid on body fat accumulation and serum lipids in hamsters fed an atherogenic diet.
Conjugated linoleic acid (CLA) refers to a mixture of naturally occurring positional and geometric isomers of linoleic acid that exist in dairy products and meat. The aim of the present work was to study the effects of c-9,t-11 and t-10,c-12 CLA isomers on body fat accumulation and serum lipids in hamsters fed an atherogenic diet. Hamsters were divided in four groups: one group was fed a chow diet (control) and the other three groups were given semi-purified atherogenic diets with 0.5% linoleic acid (LA), c-9,t-11 or t-10,c-12 CLA. Body weight and food intake were measured daily. After 6 weeks, adipose tissues from different anatomical locations and liver were dissected and weighed. Serum glucose, total cholesterol, HDL-c, LDL-c and triacylglycerol levels, as well as total and free cholesterol, triacylglycerol and phospholipid content in liver were determined by enzymatic methods. No differences in either energy intake or final body weight were found. The addition of t-10,c-12 CLA reduced fat accumulation and led to lower serum cholesterol, as compared with LA group. Nevertheless the level remained higher than in the control animals. The reduction in serum cholesterol was limited to LDL-c. This isomer also reduced triacylglycerol content in liver but did not modify serum triacylglycerol level. In summary, the present study demonstrates that t-10,c-12 CLA is the biologically active agent when anti-obesity and hypocholesterolaemic properties of CLA are considered. In contrast, the isomer c-9,t-11 has no effect on lipid metabolism in hamsters.


Management of contralateral axillary sentinel lymph nodes detected on lymphoscintigraphy for breast cancer.
Detection of a contralateral axillary sentinel lymph node (SLN) during lymphoscintigraphy for breast cancer is rare, and its significance and management are unclear. The purpose of this study was to review our experience and analyze our results together with similar patients in the literature to identify common characteristics and propose a management strategy. A PubMed search was performed for articles describing patients in whom contralateral axillary drainage was identified on lymphoscintigraphy. Additionally, a chart review was performed of all patients who had lymphoscintigraphy for breast cancer at our institution. At our institution, two of 988 (0.3 %) consecutive patients were identified with contralateral axillary drainage on lymphoscintigraphy. Twenty-seven publications describing 105 patients with contralateral axillary drainage were found. This comprised our study group of 107 patients. Lymphoscintigraphy patterns varied depending on the history and type of prior surgery. A history of chest/axillary surgery was significantly associated with absence of an ipsilateral SLN (p < 0.05). This was observed in 84.2 % of patients with prior axillary lymph node dissection versus 33.3 % with prior SLN. Contralateral SLN biopsy was attempted in 85 patients (79.4 %); 22 (20.6 %) were positive for tumor. In 17 patients (15.9 %), the contralateral node was the only positive SLN. These findings suggest that contralateral uptake on lymphoscintigraphy, though rare (0.2 %), is clinically significant and such nodes should undergo excision. Because contralateral uptake is significantly associated with prior chest/axillary surgery, routine lymphoscintigraphy should be considered in this group, as it has potential to change disease stage and management.


The effect of gamma irradiation on platelet redox state during storage.
As a method with insignificant adverse effects on in vitro quality of platelet concentrates (PCs), gamma irradiation is applied to abrogate the risk of transfusion-associated graft-vs-host disease in vulnerable recipients. However, there is some evidence of lower posttransfusion responses and proteomic alterations in gamma-irradiated platelets (PLTs), which raises some questions about their quality, safety, and efficacy. Since reactive oxygen species (ROS) are considered as markers of PLT storage lesion (PSL), the study presented here investigated oxidant state in gamma-irradiated PCs. PLT-rich plasma PC was split into two bags, one kept as control while other was subjected to gamma irradiation. Within 7 days of storage, the levels of intra-PLT superoxide, H2 O2 , mitochondrial ROS, P-selectin expression, and phosphatidylserine (PS) exposure were detected by flow cytometry while intracellular reduced glutathione (GSH), glucose concentration, and lactate dehydrogenase (LDH) activity were measured by enzymocolorimetric method. GSH decreased, while ROS generation and LDH activity increased, during storage. Gamma irradiation significantly attenuated GSH whereas increased ROS generation in earlier and later stages of storage associated with either P-selectin or PS exposure increments. Gamma irradiation can significantly increase cytosolic ROS generation in two distinct phases, one upon irradiation and another later in longer-stored PCs. While earlier ROS influx seems to be governed by direct effect of irradiation, the second phase of oxidant stress is presumably due to the storage-dependent PLT activation. Intriguingly, these observations were also in line with early P-selectin increments and increased PS exposure in longer-stored PLTs. Given the mutual link between ROS generation and PLT activation, further investigation is required to explore the effect of gamma irradiation on the induction of PSL.


When SLC46A2 delivers cargo into keratinocytes.
How pattern recognition receptors NOD1 and NOD2 sense bacterial muropeptides from extracellular bacteria to drive keratinocyte inflammation remains unclear. In this issue of Immunity, Bharadwaj et al. show that the solute carrier 46A2 (SLC46A2) delivers DAP-muropeptides into the cytosol to drive NOD1 activation in keratinocytes and elicit skin inflammation during psoriasis.


Revisiting the developmental stage and age-at-death of the "Mrs. Ples" (Sts 5) and Sts 14 specimens from Sterkfontein (South Africa): do they belong to the same individual?
During the 1947 fieldwork season, Member 4 (2-3 My) of the South African Sterkfontein site yielded two important Australopithecus africanus fossils: a cranium popularly nicknamed "Mrs. Ples" (Sts 5), and a partial skeleton (Sts 14). Previous reports have proposed that Sts 5 was a nonfully grown adolescent individual (Thackeray et al., S Afr J Sci 2002a;98:21-22), and that Sts 14 was a sub-adult specimen (according to various signs of immaturity in the skeleton) (Berge and Gommery, C R Acad Sci Paris, Sciences de la terre et des planètes 1999;329:227-232; Häusler and Berger, J Hum Evol, 2001;40:411-417; Thackeray et al., S Afr J Sci, 2002b;98:211-212). It was subsequently proposed that these fossils actually belonged to the same individual (Thackeray et al., S Afr J Sci, 2002b;98:211-212), a proposition supported by their spatial positions within the site. The present work attempts to revise these different assertions. The results obtained: (i) show that the Sts 5 fossil represents a fully grown adult cranium; (ii) provide new evidence of immaturity in the Sts 14 skeletal elements (sustaining the proposed young adult age of this specimen), and (iii) suggest that although the revised ages-at-death for these fossils are partially compatible, there is no evidence to support the idea that they represent a single individual. Finally, the encephalization quotient associated with a hypothetical union of Sts 5 and Sts 14 (calculated using data from both specimens) lies between the upper and lower limits of the currently estimated range for this species and H. habilis, respectively.


[Cannabinoid hyperemesis syndrome causing prolonged nausea and vomiting in patients with cannabis abuse].
Cannabis is one of the most used drugs worldwide. The link between repeated episodes of nausea, vomiting, and cannabis abuse is often missed in patients with prolonged cannabis abuse and is named cannabinoid hyperemesis syndrome. Characteristically, the symptoms appear in a cyclical pattern and are relieved by long, hot baths. Physical examination, radiology and endoscopy are often normal. The symptoms resolve with cessation of cannabis abuse. Health professionals must be aware of this syndrome in order to detect the patients early and to avoid extensive medical workup.


Gastric duplication cyst with ectopic pancreas in a teenager successfully resected by endoscopic submucosal dissection.
Gastric duplication cyst associated with ectopic pancreas is rare and we aimed to alert clinician to this congenital anomaly. A 15-year-old girl presented with intermittent vomiting. Gastroscopy showed a submucosal tumor with an approximate diameter of 40 mm in the anterior wall of the gastric antrum. The lesion had a central umbilication and was diagnosed preliminarily as gastric ectopic pancreas with pseudocyst formation on the basis of its appearance. However, computed tomographic scan showed a thick-walled cystic lesion with an enhanced outline of the cystic wall in the antrum of stomach, suggestive of duplication cyst. Serum amylase was normal. Endoscopic ultrasonography revealed a solid-cystic lesion; the solid portion were inhomogeneously mixed with echoes, and had indistinct border to muscularis propria; the cystic portion had echogenic internal mucosal layer and distinct border to muscularis propria. Endoscopic submucosal dissection (ESD) was suggested for the patient to relieve symptoms and diagnose the lesion definitely. The operation procedure was uneventful and the solid-cystic lesion was resected completely. Histopathologic examination revealed that the solid portion was ectopic pancreas, and the cystic portion was gastric duplication cyst. After resection, the patient discharged successfully and neither symptoms nor tumors recurred during the 9 months follow-up period. This is the first case of a solid-cystic lesion with central umbilication in the stomach diagnosed as gastric duplication cyst associated with ectopic pancreas. ESD could be an optional treatment to provide a definitive diagnosis.


Discerning the structure and energy of multiple transition states in protein folding using psi-analysis.
We quantify the degree to which folding occurs along a complex landscape with structurally distinct pathways using psi-analysis in combination with a protein engineering method that identifies native, non-covalent polypeptide interactions and their relative populations at the rate-limiting step. By probing the proximity of two specific partners, this method is extremely well-suited for comparison to theoretical simulations. Using ubiquitin as a model system, we detect individual pathways with site-resolved resolution, demonstrating that the protein folds through a native-like transition state ensemble with a common nucleus that contains heterogeneous features on its periphery. The consensus transition state topology has part of the major helix docked against four properly aligned beta-strands. However, structural heterogeneity exists in the transition state ensemble, wherein peripheral regions are differentially populated according to their relative stability. Pathway diversity reflects the variable order of formation of these peripheral elements, which radiate outward from the common nucleus. These results, which show only moderate agreement with traditional mutational phi-analysis, provide an extraordinarily detailed and quantitative description of protein folding.


Detection of biotinylated DNA probes by using Eu-labeled streptavidin and time-resolved fluorometry.
Europium has been used as a label in immunoassays as it can be measured with high sensitivity by means of time-resolved fluorometry. Here we have used streptavidin labeled with europium chelates in the detection of adenovirus type 2 DNA bound to microtiter wells after hybridization with a biotinylated probe. The method gave quantitative results and a sensitivity of about 10 pg of the specific DNA.


Spectacle Lenses With Highly Aspherical Lenslets for Slowing Myopia: A Randomized, Double-Blind, Cross-Over Clinical Trial: Parts of these data were presented as a poster at the Annual Research in Vision and Ophthalmology meeting, 2022.
To evaluate myopia progression with highly aspherical lenslet (HAL) spectacles vs conventional single vision (SV) spectacles. Prospective, double-blind, single-center, randomized, cross-over trial. A total of 119 Vietnamese children (7-13 years of age, spherical equivalent refractive error [SE] = -0.75 to -4.75D) were randomized to wear either HAL or SV, and after 6 months (stage 1) crossed over to the other lens for another 6 months (stage 2). At the end of stage 2, both groups wore HAL for a further 6 months. In the order that lenses were worn at each stage, group 1 was designated HSH (HAL-SV-HAL) and group 2 SHH (SV-HAL-HAL). The main outcome measures were a comparison between HAL and SV for change (Δ) in SE and axial length (AL) during each stage; and a comparison of ΔSE/AL with SV between HSH and SHH groups to determine whether myopia rebounded when switched from HAL to SV (HSH group). Myopia progressed more slowly with HAL than with SV during stages 1 and 2 (SEΔ stage 1: -0.21 vs -0.27D, P = .317, stage 2: -0.05 vs -0.32D, P < .001; ALΔ stage 1: 0.07 vs 0.14 mm, P = .004; stage 2: 0.04 vs 0.17 mm, P < .001). ΔSE/AL with SV was not different between the HSH and SHH groups (ΔSE -0.33 ± 0.27D vs -0.27 ± 0.42D, P = .208; ΔAL 0.17 ± 0.13mm vs 0.13 ± 0.15 mm, P = .092). An average of 14 hours per day of lens wear was reported with both lenses. In this cross-over trial, intergroup and intragroup comparisons indicate that HAL slows myopia. Children were compliant with lens wear, and data were not suggestive of rebound when patients were switched from HAL to SV.


Use of a consultant to assist a dietetic association in its media and public relations efforts.
During 1987, the Philadelphia Dietetic Association (PDA) enlisted the aid of a public relations consultant to increase media contacts and thereby accomplish a threefold goal of (a) establishing the dietetic association and its registered dietitian members as the local nutrition experts; (b) distributing reliable, current nutrition information; and (c) marketing the PDA's DIAL-A-DIETITIAN service. Through the advice of the consultant, a media tipsheet, published monthly, was created. This publication, the Nutrition Informationist, effectively increased media contacts from two in 1986 to 19 in 1987 and resulted in a total of 8 hours of air time. Weight reduction, fad diets, and sports nutrition proved to be the most popular topics. Public service announcements (PSAs) concerning the DIAL-A-DIETITIAN program were developed by the consultant to assist the public in gaining access to local registered dietitians. Calls to that service increased 75% (from an average of 47.5 to 83.0 calls) the month following the distribution of PSAs to radio stations. The positive outcome of this collaborative effort shows that public relations consultants can be a useful resource for local dietetic associations, aiding them in their efforts to reach the media and the public and to improve their image.


Prepartum transabdominal amnio-infusion for severe oligohydramnios.
Transabdominal amnio-infusion preceding labor induction was evaluated as a means of avoiding fetal distress and cesarean delivery in patients with oligohydramnios. A preliminary study was performed in 8 consecutive term or post-term pregnancies complicated by severe oligohydramnios (amniotic fluid index less than or equal to 1 cm) with unripe cervices (Bishop's score less than or equal to 3). Warm saline was injected through a spinal needle under ultrasound control. Vaginal delivery occurred in 7 cases; cesarean section was performed in one patient for failure to progress. There was no meconium aspiration and no sign of fetal distress.


Tricuspid insufficiency does not increase early after permanent implantation of pacemaker leads.
Interference between pacemaker (PM) lead and tricuspid apparatus may cause tricuspid regurgitation (TR). However, data regarding TR in patients with implanted PM are controversial. Our aim is to find out the degree of TR in a group of patients before and following PM implantation in a prospective manner. The study group consisted of the patients referred for implantation of permanent PM or implantable cardioverter defibrillator (ICD). All patients underwent two-dimensional and Doppler echocardiographic evaluation before and after device implantation. The severity of TR was qualitatively classified into four groups as normal or trivial, mild, moderate, or severe. All studies were reviewed for accuracy by a second independent interpreter. Sixty-one patients (mean age 53 +/- 8 years, 44 male) referred for PM (n = 55) or ICD (n = 6) implantation consisted of the study population. Echocardiographic degree of TR was mild in 21 (70%), moderate in 7 (23%) and severe in 2 (7%) patients before PM implantation. Following device implantation, mild TR was noted in 23 (76%), moderate in 10 (33%), and severe in 2 (6%) cases. After the procedure, the TR severity was increased from normal/trivial to mild in 5 (16%) cases and from mild to moderate in 3 (10%). There was no worsening of the severity of TR in patients with moderate regurgitation following device implantation. The severity of TR did not change at a mean follow-up of 6 +/- 3 months. New or worsening TR is relatively rare after PM implantation. It is not associated with an acute worsening or clinical deterioration. But echocardiographic follow-up is recommended to monitor other complications in chronic phase.


Flocculation as a low-cost method for harvesting microalgae for bulk biomass production.
The global demand for biomass for food, feed, biofuels, and chemical production is expected to increase in the coming decades. Microalgae are a promising new source of biomass that may complement agricultural crops. Production of microalgae has so far been limited to high-value applications. In order to realize large-scale production of microalgae biomass for low-value applications, new low-cost technologies are needed to produce and process microalgae. A major challenge lies in the harvesting of the microalgae, which requires the separation of a low amount of biomass consisting of small individual cells from a large volume of culture medium. Flocculation is seen as a promising low-cost harvesting method. Here, we overview the challenges and possible solutions for flocculating microalgae.


Living with death: the evolution of the mitochondrial pathway of apoptosis in animals.
The mitochondrial pathway of cell death, in which apoptosis proceeds following mitochondrial outer membrane permeabilization, release of cytochrome c, and APAF-1 apoptosome-mediated caspase activation, represents the major pathway of physiological apoptosis in vertebrates. However, the well-characterized apoptotic pathways of the invertebrates C. elegans and D. melanogaster indicate that this apoptotic pathway is not universally conserved among animals. This review will compare the role of the mitochondria in the apoptotic programs of mammals, nematodes, and flies, and will survey our knowledge of the apoptotic pathways of other, less familiar model organisms in an effort to explore the evolutionary origins of the mitochondrial pathway of apoptosis.


Protein backbone motions viewed by intraresidue and sequential HN-Halpha residual dipolar couplings.
Triple resonance E.COSY-based techniques were used to measure intra-residue and sequential H(N)-H(alpha) residual dipolar couplings (RDCs) for the third IgG-binding domain of protein G (GB3), aligned in Pf1 medium. Measurements closely correlate with values predicted on the basis of an NMR structure, previously determined on the basis of a large number of one-bond backbone RDCs measured in five alignment media. However, in particular the sequential H(N)-H(alpha) RDCs are smaller than predicted for a static structure, suggesting a degree of motion for these internuclear vectors that exceeds that of the backbone amide N-H vectors. Of all experimentally determined GB3 structures available, the best correlation between experimental (1)H-(1)H couplings is observed for a GB3 ensemble, previously derived to generate a realistic picture of the conformational space sampled by GB3 (Clore and Schwieters, J Mol Biol 355:879-886, 2006). However, for both NMR and X-ray-derived structures the (1)H-(1)H couplings are found to be systematically smaller than expected on the basis of alignment tensors derived from (15)N-(1)H amide RDCs, assuming librationally corrected N-H bond lengths of 1.041 A.


Large neutral amino acid status in association with P:T ratio and diet in adult and pediatric patients with phenylketonuria.
Intake of large neutral amino acids (LNAA) may inhibit phenylalanine (PHE) transport across the blood brain barrier and assist with blood PHE control in patients with phenylketonuria (PKU). We evaluated the interrelationship between LNAA in plasma and diet on Phe:Tyr (P:T) ratio in patients with PKU and the influence of dietary factors on plasma LNAA markers. Plasma amino acid values and 3-day food record analysis from two studies (34 male/30 female; age 4.6-47 years) were examined. For pediatrics (<18 years) and adults (≥18 years) the relationship between P:T ratio, plasma LNAA, and dietary intake patterns were investigated. Dietary factors influencing P:T ratio included intake of total protein (g/kg), medical food (MF) protein (g/kg, % below Rx), and LNAA (g) in the full cohort (P < .05). Associations were found between plasma valine and other dietary and plasma LNAA in pediatrics (P < .05) and plasma LNAA with dietary LNAA intake in adults (P = .019). Plasma P:T ratio was inversely associated with plasma LNAA concentrations in both age groups (P < .05). Aside from histidine in pediatrics (P = .024), plasma LNAA did not differ by having plasma PHE levels within or above the therapeutic range (120-360 μmol/L). Plasma LNAA in both age groups was similar to reported healthy control values. P:T ratio is significantly tied to dietary LNAA, adherence to MF Rx, and plasma LNAA concentrations. Additionally, P:T ratio and valine may be effective clinical proxies for determining LNAA metabolic balance and LNAA quality of the diet in patients with PKU.


Response of hepatoma 9618a and normal liver to host carbogen and carbon monoxide breathing.
The effects of hyperoxia (induced by host carbogen [95% oxygen/5% carbon dioxide breathing] and hypoxia (induced by host carbon monoxide [CO at 660 ppm] breathing) were compared by using noninvasive magnetic resonance (MR) methods to gain simultaneous information on blood flow/oxygenation and the bioenergetic status of rat Morris H9618a hepatomas. Both carbogen and CO breathing induced a 1.5- to 2-fold increase in signal intensity in blood oxygenation level dependent (BOLD) MR images. This was due to a decrease in deoxyhemoglobin (deoxyHb), which acts as an endogenous contrast agent, caused either by formation of oxyhemoglobin in the case of carbogen breathing, or carboxyhemoglobin with CO breathing. The results were confirmed by observation of similar changes in deoxyHb in arterial blood samples examined ex vivo after carbogen or CO breathing. There was no change in nucleoside triphosphates (NTP)/P(i) in either tumor or liver after CO breathing, whereas NTP/P(i) increased twofold in the hepatoma (but not in the liver) after carbogen breathing. No changes in tumor intracellular pH were seen after either treatment, whereas extracellular pH became more alkaline after CO breathing and more acid after carbogen breathing, respectively. This tumor type and the liver are unaffected by CO breathing at 660 ppm, which implies an adequate oxygen supply.


Economization of cerebral activation under training: The inverse U- shaped function revisited.
As already observed as early as 1967 by Ingvar and Risberg in their pioneering work, effects of practice of working memory on cerebral functions have been confirmed repeatedly in a number of neuroimaging studies using positron emission tomography (PET) and functional magnetic resonance imaging (fMRI). Accordingly, initial performance gains are typically accompanied by increased cerebral activation, while consolidation of such performance gains goes along with a subsequent decrease in activation resembling an inverse U-shaped function. This observed pattern can be interpreted as an economization of cerebral functioning as tasks are being accomplished with relatively lower effort and may also apply to other cognitive domains. However, the economization of cerebral activation under training may depend on task difficulty and training characteristics on the one hand and individual factors, including age, intelligence, cognitive reserve, education, physical and mental health on the other. These findings bear important implications for the design of neuroimaging studies and stimulation protocols, in which similar tasks are routinely repeated.


Effect of Surgical Treatment for N2-Positive c-stage III Non-Small Cell Lung Carcinoma in the "PACIFIC" Era.
The PACIFIC trial findings drastically changed the c-stage III non-small cell lung cancer (NSCLC) treatment strategy. However, it remains uncertain whether surgery is no longer needed for treatment. We aimed to evaluate the efficacy of surgery and explore the prognostic factors of better outcomes in surgery-treated patients than in PACIFIC regimen-treated patients. From 2010 to 2020, 107 patients with clinical N2-stage III NSCLC underwent lung resection in our institute. We analyzed and compared the yearly postoperative overall survival (OS) benchmarks of these patients to those of patients treated in the PACIFIC trial. The 1-, 2-, 3-, 4-, and 5-year OS rates of patients were 87.7%, 73.9%, 64.9%, 58.2%, and 55.4%, respectively, all of which were superior to those of PACIFIC regimen-treated patients. However, patients with cT3/T4 tumors and skip, multistation, distant, and bulky N2 metastases, as well as those who underwent bronchoplasty, showed inferior results in several yearly benchmarks than in PACIFIC regimen-treated patients. Multivariate analyses conducted among factors mentioned above showed that cT3/T4 tumor was a worse prognostic factor for surgery-treated patients than for PACIFIC regimen-treated patients (hazard ratio [HR] 1.89, P = .036). Distant N2 metastasis was also a worse prognostic factor, although its effect was not statistically significant (HR 1.81, P = .082). Surgery remains the mainstay of N2-positive c-stage III NSCLC treatment, and the PACIFIC regimen may be suitable only for patients with unresectable disease. However, surgery should be cautiously considered for patients with cT3/4 disease.


Exploring the epigenetic regulation of telomerase reverse transcriptase (TERT) in human cancer cell lines.
Telomerase regulation, including TERT promoter methylation, has been of long-standing interest to cancer biologists. Rowland et al. have now vastly expanded their ongoing characterization of TERT promoter methylation in cancer cells, analyzing the methylation patterns of 833 cell lines from 23 human cancers. They document a highly conserved pattern of hypomethylation around the proximal promoter, as well as a more heterogeneous region of hypermethylation further upstream, both associated with active TERT expression in cancer cells. They further describe the interplay between activating TERT promoter mutations and allelic methylation and transcription patterns. This valuable dataset represents the most extensive characterization of TERT promoter methylation in cancer cells to date and will help guide the future study of transcriptional regulation of telomerase. Comment on: https://doi.org/10.1002/1878-0261.12786.


Physiopathologic aspects of ileal reservoirs.
The author reviews some aspects of fecal continence after ileoanal anastomosis with an ileal reservoir. Also, the consequences of fecal stasis on small-bowel absorption and morphologic and bacteriologic features are discussed. Some recommendations for the follow-up of these patients are outlined.


A PCA approach to population analysis: with application to a Phase II depression trial.
For psychiatric diseases, established mechanistic models are lacking and alternative empirical mathematical structures are usually explored by a trial-and-error procedure. To address this problem, one of the most promising approaches is an automated model-free technique that extracts the model structure directly from the statistical properties of the data. In this paper, a linear-in-parameter modelling approach is developed based on principal component analysis (PCA). The model complexity, i.e. the number of components entering the PCA-based model, is selected by either cross-validation or Mallows' Cp criterion. This new approach has been validated on both simulated and clinical data taken from a Phase II depression trial. Simulated datasets are generated through three parametric models: Weibull, Inverse Bateman and Weibull-and-Linear. In particular, concerning simulated datasets, it is found that the PCA approach compares very favourably with some of the popular parametric models used for analyzing data collected during psychiatric trials. Furthermore, the proposed method performs well on the experimental data. This approach can be useful whenever a mechanistic modelling procedure cannot be pursued. Moreover, it could support subsequent semi-mechanistic model building.


Clinical research progress on BRAF V600E-mutant advanced colorectal cancer.
Colorectal cancer is one of the malignant tumors that pose a serious threat to human health. A particularly bad prognosis might be expected for colorectal tumors with the unique molecular subtype BRAF V600E mutation. With the development of precision therapy, the advent of molecularly targeted therapies and immune checkpoint inhibitors has improved the outcome of intermediate to advanced colorectal cancer. However, the duration of drug benefit is usually short, and overall survival and progression-free survival remain suboptimal. Therefore, investigators are exploring more rational, safe, and effective drug combination regimens through clinical trials to provide longer survival for patients with such genetic mutations with metastatic colorectal cancer (mCRC). This article reviews the progress of clinical research on molecularly targeted drugs, immune checkpoint inhibitors, first-line chemotherapeutic agents, and different combination therapy regimens (including different targeted drug combinations, immune combination targeting, and chemotherapy combination targeting) for colorectal cancer patients with BRAF V600E mutation, which provides a reference for further in-depth clinical exploration of the treatment of colorectal cancer patients with BRAF V600E mutation.


Highlights from the 12th congress of the international pediatric transplant association, Austin, Texas 2023.
The 12th Congress of the (IPTA) event in Austin, Texas, had over 400 attendees from 40 countries. The attendees included a diverse mix of pediatric transplant professionals from several specialties including physicians, surgeons, scientists, nurses, organ procurement personnel, advance transplant providers, pharmacists, administrators, fellows, residents, and students. The 4-day event featured nearly 200 abstracts, 90 oral presentations, 24 mini oral presentations, and more than 80 poster presentations. All of these presentations encouraged vibrant discussions and supported the exchange of new clinical and basic science information regarding clinical care management, basic science research, socioeconomic, and ethical and organ donation issues relevant to pediatric transplantation. We briefly describe here the highest scored presented abstracts at IPTA 2023 that are divided into two categories: clinical and basic sciences.


Crystal structure of 4,10-dimeth-oxy-13-methyl-6H,12H-6,12-epimino-dibenzo[b,f][1,5]dioxocine.
The title compound, C17H17NO4, lacks crystallographic symmetry with one mol-ecule per asymmetric unit. The mol-ecule exists in a folded butterfly-like conformation; the benzene rings form a dihedral angle of 84.72 (7)°. The central eight-membered imino-bridged dioxocin ring adopts a twisted-boat conformation. In the crystal, inversion-related mol-ecules are linked by pairs of weak C-H⋯O hydrogen bonds, forming double-stranded chains parallel to the a axis.


How international are leading general paediatric journals?
The composition of the editorial boards of leading general paediatric journals was assessed. The board members' country of current affiliation was classified according to the Human Development Index. The results showed that only a very small number of the board members are based in the developing world.


Interactive effects of traits, states, and gender on cardiovascular reactivity during different situations.
Interactive effects of anger and anxiety traits, negative affect state, different situations, and gender on cardiovascular reactivity (CVR) to stress were examined. Subjects (91 men, 92 women) performed a reaction time task under either a Social Evaluation, a Harassment, or a Control condition; SBP, DBP, and HR were recorded continuously. Hierarchical multiple regressions revealed intricate interactions. The interaction of anger expression style and anger experience was significant only among men, such that anger suppressors with high trait anger showed the largest CVR of any group during Harassment; anger expressors exhibited generally high CVR across conditions. However, anger expression style and state negative affect interacted to affect CVR in both men and women. Finally, the fear of negative evaluation predicted elevated DBP responses only among men in the Social Evaluation condition. Results imply that the extent to which traits of anger and anxiety contribute to coronary risk may depend on interactions with other traits, gender, and the environment.


Personality development following sexual abuse.
Thirty-seven girls and twelve boys who had been sexually abused at an average of 2.6 years previously were traced and reviewed. The nonoffending parents participated in a structured interview and the children were assessed using the Piers-Harris Self-Concept Scale and the Achenbach Child Behavior Checklist. Each child was matched with a child not known to have been sexually abused and these control children underwent a similar psychological assessment. Interviews with the nonoffending parents found that 76% of the children were thought to be less confident than before, 30% had fewer friends, and 20% were more aggressive. Increased sexual awareness was noted in 24%. School teachers reported that 28% still had behavior problems, 17% had repeated a year at school, and a further 17% had deteriorated in their school work. The sexually abused girls had significantly lower self-esteem than the control girls. There was no difference in self-esteem between the control and the sexually abused boys. There was a higher incidence of stranger assault in the boys compared with the girls. The Child Behavior Checklist completed by the parents, the Teacher Report Form, and the Youth Self-Report of the Child Behavior Checklist showed that a highly significant proportion of sexually abused children fell into the clinical range. Sexual abuse appears to have long-term adverse consequences for many of the victims. This may have implications for their ability to relate to others, for the adult friendships they will make, and eventually for the way they will relate to their own children.


Molecular diagnosis of clear cell sarcoma: detection of EWS-ATF1 and MITF-M transcripts and histopathological and ultrastructural analysis of 12 cases.
Clear cell sarcoma (CCS), also known as melanoma of soft parts, is an uncommon deep soft tissue tumor presenting typically in the lower extremities of young adults. Previous cytogenetic studies have established the specificity of the recurrent t(12;22)(q13;q12), resulting in a EWS-ATF1 fusion, for CCS. The prevalence of the EWS-ATF1 fusion in CCS remains unclear, since most genetically confirmed CCS have been reported as isolated cytogenetic or molecular diagnostic case reports. We therefore studied histologically confirmed CCS from 12 patients for the presence of EWS-ATF1 by reverse-transcriptase polymerase chain reaction (RT-PCR), using RNA extracted from either frozen (four cases) or formalin-fixed paraffin-embedded (eight cases) material. All primary tumors were located in the deep soft tissues of the extremities. Histologically, 10 cases had a typical epithelioid nested appearance. Most or all cases showed immunostaining for HMB45 (12 of 12), S-100 protein (10 of 12), and MITF (12 of 12). Ultrastructural analysis showed melanosomes in six of seven cases. The presence of an EWS-ATF1 fusion transcript was identified by RT-PCR in 11 of 12 cases (91%), all of which showed the same fusion transcript structure, namely the previously described in-frame fusion of EWS exon 8 to ATF1 codon 65. RT-PCR analysis for the melanocyte-specific splice form of the MITF transcript was positive in all cases tested (4 of 4). These data confirm that EWS-ATF1 detection can be used as a highly sensitive diagnostic test for CCS and that CCS expresses the melanocyte-specific form of the MITF transcript, further supporting its genuine melanocytic differentiation.


A new approach for two-terminal electronic memory devices - Storing information on silicon nanowires.
The work described in this paper focuses on the utilisation of silicon nanowires as the information storage element in flash-type memory devices. Silicon nanostructures have attracted attention due to interesting electrical and optical properties, and their potential integration into electronic devices. A detailed investigation of the suitability of silicon nanowires as the charge storage medium in two-terminal non-volatile memory devices are presented in this report. The deposition of the silicon nanostructures was carried out at low temperatures (less than 400 °C) using a previously developed a novel method within our research group. Two-terminal non-volatile (2TNV) memory devices and metal-insulator-semiconductor (MIS) structures containing the silicon nanowires were fabricated and an in-depth study of their characteristics was carried out using current-voltage and capacitance techniques.


Quantification of low levels of rainbow trout immunoglobulin by enzyme immunoassay using two monoclonal antibodies.
An enzyme immunoassay has been developed for quantitation of low levels of trout immunoglobulin (Ig). This assay uses two monoclonal antibodies, one as capture antibody and the other as detector, directed against two non-overlapping epitopes on the heavy chains of trout Ig. The assay shows high reproducibility and can detect 0.12 micrograms trout Ig ml-1. Coefficients of intra- and interassay variation ranged from 3.8 to 7.1% and from 7.9 to 17.4%, respectively. Analysis of 37 healthy trout showed increasing serum Ig concentration with size. The mean Ig concentration was 0.67 mg ml-1 for trout of about 20 g and 9.1 mg ml-1 for trout weighing more than 125 g.


Venous thromboembolism in immobilized patients with dementia. Findings from the RIETE registry.
The natural history of venous thromboembolism (VTE) in patients with dementia has not been thoroughly studied. We used the RIETE Registry data to assess the clinical characteristics, treatment strategies and outcome during the first 3 months after acute VTE in all immobilized patients with dementia. As of August 2011, 37988 patients had been enrolled, of whom 1316 (3.5%) had dementia. Most patients in both subgroups were initially treated with low-molecular-weight heparin (LMWH). Then, 48% of patients with dementia and 25% of those without dementia received LMWH as long-term therapy. During the first 3 months of anticoagulant therapy, patients with dementia had a higher incidence of fatal pulmonary embolism (PE): 4.0% vs. 1.2% (odds ratio: 3.3; 95% CI: 2.5-4.4) and fatal bleeding: 1.4% vs. 0.5% (odds ratio: 2.9; 95% CI: 1.8-4.6) than those without dementia. In demented patients initially presenting with PE, the incidence of fatal PE during the first week outweighed that of fatal bleeding (42 vs. 4 deaths), but from Day 8, the incidence of fatal PE was similar to the incidence of fatal bleeding. In patients initially presenting with deep vein thrombosis (DVT), there were 4 fatal PE and 8 fatal bleeding events. VTE patients with dementia had a high incidence of fatal PE and fatal bleeding. In those initially presenting with PE, the risk of dying of PE far outweighed that of fatal bleeding. In patients presenting with DVT alone, the risk of fatal PE was lower than that of fatal bleeding.


Emulsified perfluorochemicals for oxygenation of microbial cell cultures?
PFCs and their emulsions may have value for increasing the efficiency of O2-transport to microbial cultures. Therefore, the effects of emulsion components on growth of S. cerevisiae and E. coli have been examined. Viable cell counts revealed that perfluorodecalin or the commercial emulsion, Fluosol-DA 20%, produced no obvious growth-inhibition over 6h. However, incubation of cells with up to 10% of the Pluronic F-68 surfactant reduced absorbance at 600nm. Further experiments to assess the effects of PFC emulsion components on growth and structure of microbial cells are in progress.


Retrieving orbital angular momentum distribution of light with plasmonic vortex lens.
We utilize a plasmonic vortex lens (PVL) to retrieve the orbital angular momentum (OAM) distribution of light. The OAM modes are coupled to the surface plasmon polaritons (SPPs) in the form of various Bessel functions respectively. By decomposing the interference pattern of SPPs into these Bessel functions, we can retrieve the relative amplitude and the relative phase of input OAM modes simultaneously. Our scheme shows advantage in integration and can measure hybrid OAM states by one measurement.


Myeloid Cells Restrict MCMV and Drive Stress-Induced Extramedullary Hematopoiesis through STAT1.
Cytomegalovirus (CMV) has a high prevalence worldwide, is often fatal for immunocompromised patients, and causes bone marrow suppression. Deficiency of signal transducer and activator of transcription 1 (STAT1) results in severely impaired antiviral immunity. We have used cell-type restricted deletion of Stat1 to determine the importance of myeloid cell activity for the defense against murine CMV (MCMV). We show that myeloid STAT1 limits MCMV burden and infection-associated pathology in the spleen but does not affect ultimate clearance of infection. Unexpectedly, we found an essential role of myeloid STAT1 in the induction of extramedullary hematopoiesis (EMH). The EMH-promoting function of STAT1 was not restricted to MCMV infection but was also observed during CpG oligodeoxynucleotide-induced sterile inflammation. Collectively, we provide genetic evidence that signaling through STAT1 in myeloid cells is required to restrict MCMV at early time points post-infection and to induce compensatory hematopoiesis in the spleen.


Synthesis and antimicrobial activities of novel naphtho[2,1-b]pyran, pyrano[2,3-d]pyrimidine and pyrano[3,2-e][1,2,4]triazolo[2,3-c]-pyrimidine derivatives.
The synthesis of new naphtho[1',2':5,6]pyrano[2,3-d]pyrimidines and related heterocycles has been reported. The key intermediate 3-amino-8-bromo-1-(p-methoxyphenyl)-1H-naphtho[2,1-b] pyran-2-carbonitrile (3c) was obtained in one pot synthesis by treating alpha-cyanocinnamonitrile (1c) with 6-bromo-2-naphthol (2). Antimicrobial activity was shown for some of the synthesized compounds.


Prevalence of Helicobacter pylori resistance to metronidazole, clarithromycin, amoxycillin and tetracycline in Lebanon.
The aim of this study was to determine the prevalence of antimicrobial resistance amongst Helicobacter pylori isolates cultured from gastric biopsies taken during routine endoscopies at the American University of Beirut Medical Center in Lebanon. Fifty-four consecutively recovered H. pylori isolates were tested against metronidazole, clarithromycin, tetracycline and amoxycillin using the epsilometer test. Resistance to metronidazole (MIC > 8 mg/l) was found in 29.5% of the isolates while resistance to clarithromycin (MIC > or =0.25 mg/l) and tetracycline (MIC > or =4 mg/l) was low (4 and 2%, respectively). All isolates were susceptible to amoxycillin (MIC < or =8 mg/l). These findings are comparable with those reported from Europe and the United States. The prevalence of metronidazole resistance in our study was lower than that from other parts of the Middle East and the developing world.


Dilute prothrombin time-based lupus ratio test. Integrated LA testing with recombinant tissue thromboplastin.
The lupus ratio (LR) test is a normalized ratio of the clotting times obtained with low and high phospholipid (PL) concentrations, where the test plasma is mixed 1:1 with normal pooled plasma (NP). As an integrated, automated, and computer-assisted assay, this principle has been applied to the dilute activated partial thromboplastin time (dAPTT) and dilute Russell viper venom time (dRVVT) test systems. In this study, we used recombinant thromboplastin to develop an automated LR test based on the dilute prothrombin time (dPT). Using plasma samples from a selected group of patients (N=92) with a well-defined lupus anticoagulant (LA) status, the dPT-based LR test showed fair agreement with the dAPTT-based LR test (kappa=.60, P<.001) and good agreement with the dRVVT-based LR test (kappa=.78, P<.001) regarding the classification of plasmas as LA-negative or -positive. Most discordant plasmas were low positive. Correlation between dPT- and dRVVT-positive LRs was only moderate (r(s)=.51, P=.002), and correlation between dPT- and dAPTT-positive LRs was nonsignificant (r(s)=.23, P=.18). Using dilutions of pooled LA-positive plasma, the dAPTT-based LR test appeared to be the most sensitive of the three tests. This study confirms the general validity of the LR principle and shows that the dPT-based LR test yields reproducible results. The low coefficients of variation (CV) of the LR assays, combined with a fairly low correlation among the three tests, support the assumption of antibody heterogeneity of LA-positive plasmas and the desirability of performing more than one test.


The frequency of glove contamination during cesarean delivery.
To determine the frequency of glove contamination associated with fetal extraction during cesarean delivery. The study was performed in 25 women having scheduled or unscheduled cesarean delivery. Surgeons double-gloved for all procedures. Immediately before and after delivery of the fetus, the dorsal aspect of the fingers and hand of the surgeon's outer glove was swabbed with cotton-tip applicators and cultured for aerobic and anaerobic organisms. Only the glove from the hand that was used to deliver the infant was cultured. Nine of 25 cultures (36%, 95% confidence interval [CI] 17-55) performed immediately before fetal extraction were positive for staphylococci. No other organisms were isolated. Cultures performed following fetal extraction showed non-staphylococcal bacteria in 11 of 14 (79%, 95% CI 58-100) laboring women and one of 11 (9%, 95% CI 0-26) nonlaboring women, a statistically significant difference (P < .01). In the laboring patients, non-staphylococcal bacteria were isolated with similar frequency from the dorsal aspect of the hand (seven of 14, 50%, 95% CI 24-76) and the fingers (ten of 14, 71%, 95% CI 47-95). These cultures yielded mostly bacterial species from the Enterobacteriaceae family. In laboring patients with ruptured membranes, delivery of the fetal head frequently results in contamination of the surgeon's glove with pathogenic bacteria. This finding may partially explain the increased frequency of post-cesarean endometritis associated with manual extraction of the placenta.


Effect of digitonin on membrane-bound and chitosomal chitin synthetase activity in protoplasts from yeast cells of Candida albicans.
The effect of digitonin on chitin synthetase present in membrane (MMF) and cytoplasmic fractions (chitosomes) (CF) from C. albicans yeast protoplasts has been determined. The zymogen is preferentially, but not exclusively, solubilized by digitonin from MMF. Centrifugation of distinct solubilized preparations, containing either zymogen, in vivo active enzyme and/or trypsin activated enzyme, on linear sucrose gradients suggests that both zymogen and trypsin activated enzyme sediment slightly slower than the active enzyme, pointing out differences between the activation processes in vivo and in vitro or, alternatively, that both enzyme activities (active in vivo and zymogenic) correspond to different gene products. The detection of a zymogenic activity under certain conditions (0.5 mg ml-1 of digitonin and 64 micrograms ml-1 of trypsin) also suggests the existence of more than one pool of zymogenic enzyme in the MMF. Digitonin sensitizes the chitosomal (CF) proenzyme to trypsin: activation is enhanced by low digitonin concentrations in the presence of 8 micrograms ml-1 of protease, whereas activity strongly decreases in the presence of 64 micrograms ml-1 of trypsin. Digitonin does not produce zymogen activation per se in absence of exogenous protease. Furthermore, chitosome structure is modified into particles with low buoyant densities.


Reduction of the dose to the elective neck in head and neck squamous cell carcinoma, a randomized clinical trial using intensity modulated radiotherapy (IMRT). Dosimetrical analysis and effect on acute toxicity.
A randomized trial was initiated to investigate whether a reduction of the dose to the elective nodal sites and the swallowing apparatus delivered by IMRT would result in a reduction of acute and late side effects without compromising tumor control. The aim of this paper is to report on dosimetrical analysis and acute toxicity. Two-hundred patients were randomized. In the standard arm, elective nodal volumes (PTVelect) were irradiated up to an equivalent dose of 50Gy. In the experimental arm an equivalent dose of 40Gy was prescribed to the PTVelect. The dose to the swallowing apparatus was kept as low as possible without compromising therapeutic PTV (PTVther) coverage. No significant difference was seen between both arms concerning PTVther coverage. The median D95 of the PTVelect was significantly lower in the experimental arm (39.5 vs 49.8Gy; p<0.001). Concerning the organs at risk, the dose to swallowing structures and spinal cord was significantly reduced. There was no significant difference in acute toxicity. Three months after radiotherapy there was significantly less grade ⩾3 dysphagia in the experimental arm (2% vs 11%; p=0.03). With a median follow-up of 6months no significant differences were observed in locoregional control, disease free survival or overall survival. Using IMRT we were able to significantly reduce the dose to the PTVelect and several organs at risk without compromising PTVther coverage. This resulted in a significant reduction of severe dysphagia 3months after radiotherapy. Further follow-up is necessary to investigate whether these observations translate into a benefit on late treatment related dysphagia without affecting treatment outcome.


Differences in XPS and solid state NMR spectral data and thermo-chemical properties of iso-structural compounds in the series KTaF6, K2TaF7 and K3TaF8 and KNbF6, K2NbF7 and K3NbF8.
A series of compounds KTaF6, K2TaF7 and K3TaF8 and KNbF6, K2NbF7 and K3NbF8 was investigated by means of XPS and MAS NMR spectroscopy and DSC measurements. Electron binding energies of all accessible orbitals were discussed and, for the first time, correlations between different orbital energies were examined. (19)F MAS NMR data and other NMR parameters of the investigated compounds were correlated with structural information, as well as with XPS data. Also a complete set of DSC data was presented including a number of phase transitions and their heat contents. Based on measured characteristics it was shown how differences in the electronic structure of isostructural compounds influence the spectral and thermo-chemical behaviour of the investigated pairs, i.e. KTaF6vs. KNbF6, K2TaF7vs. K2NbF7 and K3TaF8vs. K3NbF8. It was concluded that the differences in K 2s orbital energies play an important role in the different behaviour of tantalate and niobate analogues.


The effects of platelet-derived growth factor in rat cardiac allograft vasculopathy and fibrosis.
We investigated the relationship between expression of platelet-derived growth factor (PDGF) and development of chronic rejection. Rat recipients pretreated by inoculation of donor splenocytes (SPC) followed by cyclophosphamide (CP) were divided into three groups: control animals without immunosuppression (group 1, n = 10); group 2 recipients treated with cyclosporine (n = 10), and euthanized at 2 to 3 months posttransplantation; group 3 (n = 20) animals treated with SPC and CP with 10 recipients euthanized at 2 weeks posttransplantation and 10 monitored for at least 1 year. Immunohistological and pathological studies were performed to evaluate cardiac allograft vasculopathy (CAV), myofibrosis, and expression of PDGF. Allografts collected from group 2 demonstrated abundant infiltration of collagenous fibers in the coronary arteries. Edema and proliferation of the intima as well as the lumial stenosis were revealed in groups 1 and 2. Additionally, immunohistological staining demonstrated a high expression of PDGF protein in cardiac muscles and coronary arteries in groups 1 and 2. In contrast, pretreatment of animals with SPC plus CP induced long-term allograft survival. Importantly, the expression of PDGF in cardiac muscles and arteries in group 3 was significantly less than groups 1 and 2. CAV were dramatically reduced in group 3 when compared with groups 1 and 2. Preconditioning of recipients with SPC and CP suppressed the expression of PDGF within allografts and induced transplant tolerance. Our results suggested that the expression of PDGF plays an important role during the development of cardiac allograft rejection.


Skin microbiota is the main reservoir of Roseomonas mucosa, an emerging opportunistic pathogen so far assumed to be environmental.
Roseomonas spp. are increasingly involved in human infectious diseases. The environmental source for infection is generally admitted in published cases owing to the origin of most Roseomonas species and to their affiliation to the family Acetobacteraceae in Rhodospirillales, which mainly groups environmental bacteria. For a better delineation of Roseomonas habitat and infectious reservoir, we related phenotype, phylotype (16S rRNA gene), genomotype (pulsed-field gel electrophoresis) and origin of 33 strains isolated from humans, hospital environment and natural environment. Genetic and metagenomic databases were also surveyed. The population structure of the genus showed clades associated with humans, whereas others grouped environmental strains only. Roseomonas mucosa is the main human-associated species and the study supported the idea that opportunistic infections due to this species are related to the patient skin microbiota rather than to the environment. In contrast, some strains belonging to other species isolated from patients with cystic fibrosis were related to environmental clades, suggesting an exogenous source for patient colonization. Accurate knowledge about the reservoirs of opportunistic pathogens that have long been considered of environmental origin is still needed and would be helpful to improve infection control and epidemiological survey of emerging human pathogens.