Cell migration and cortisone induction of sucrase activity in jejunum and ileum.
The increase of sucrase activity in homogenates of jejunum and ileum of suckling rats after cortisone administration has been investigated. Serial tissue sections of villi and crypts were also assayed for sucrase activity and these results were compared with the migration of cells labelled with [(3)H]thymidine along the villus. By using a low dose of cortisone (0.5mg/day per 100g body wt.) it was found that the sensitivity of the small intestine producing system to cortisone stimulation increased during the suckling period. On the other hand, 5mg of cortisone/day per 100g body wt. produced practically the same increase of sucrase during the entire suckling period. Sucrase activity in homogenates of the entire small-intestinal wall was first detected 24h after the first injection of cortisone (5mg/day per 100g body weight) to 9-day-old animals and maximum activity both in the jejunum and ileum was reached by 120h. Jejunal activity was greater than ileal activity, but the rate of the increase was similar. The half-time of the increase was 23-27h, whereas enterocytes migrate from the base to the tip of the villi in approximately 72h. Comparison of sucrase activity in serial tissue sections of villi and crypts at various times after cortisone treatment showed that the leading edge of sucrase activity proceeds toward the tip of the villi at the same rate as the advancing edge of newly formed cells. Sucrase activity increased in the newly induced cells as they migrated to the tip of the villi. It was concluded that the increase of sucrase activity in suckling rats after cortisone stimulation is due to at least three factors: (1) increase of activity in newly differentiating cells, (2) increased percentage of villus cells with sucrase activity and (3) continued production or activation of sucrase activity as the cells migrate along the villi.


The insecticidal neurotoxin Aps III is an atypical knottin peptide that potently blocks insect voltage-gated sodium channels.
One of the most potent insecticidal venom peptides described to date is Aps III from the venom of the trapdoor spider Apomastus schlingeri. Aps III is highly neurotoxic to lepidopteran crop pests, making it a promising candidate for bioinsecticide development. However, its disulfide-connectivity, three-dimensional structure, and mode of action have not been determined. Here we show that recombinant Aps III (rAps III) is an atypical knottin peptide; three of the disulfide bridges form a classical inhibitor cystine knot motif while the fourth disulfide acts as a molecular staple that restricts the flexibility of an unusually large β hairpin loop that often houses the pharmacophore in this class of toxins. We demonstrate that the irreversible paralysis induced in insects by rAps III results from a potent block of insect voltage-gated sodium channels. Channel block by rAps III is voltage-independent insofar as it occurs without significant alteration in the voltage-dependence of channel activation or steady-state inactivation. Thus, rAps III appears to be a pore blocker that plugs the outer vestibule of insect voltage-gated sodium channels. This mechanism of action contrasts strikingly with virtually all other sodium channel modulators isolated from spider venoms that act as gating modifiers by interacting with one or more of the four voltage-sensing domains of the channel.


Performance analysis of 8-channel MDCT angiography in detection, localization, and sizing of intracranial aneurysms identified on DSA.
To retrospectively analyze the performance of 8-channel multi-detector computed tomographic angiography (MDCTA) in detecting, localizing, and sizing intracranial aneurysms using conventional intraarterial digital subtraction angiography (DSA) as the gold standard. A retrospective search from November 2004 to August 2006 revealed subarachnoid hemorrhage (SAH) in 25 patients (13 females and 12 males, age range 21-73 years) who underwent both DSA and 8-channel MDCTA exams. Two independent reviewers compared both studies for the presence, location, and size of the aneurysm. A total of 35 aneurysms in 25 patients were identified on DSA, and 33 on MDCTA. MDCTA missed two 4-mm posterior communicating artery (PcoA) aneurysms. There was a mismatch in the location of two aneurysms. One aneurysm at PcoA on DSA was reported as a supraclinoid internal carotid artery (ICA) aneurysm on MDCTA. The other on M2 bifurcation was reported as on M1. No size mismatch was noted. MDCTA had a 94% (33/35) aneurysm detection rate, 88% (31/35) aneurysm localization rate and 100% (33/33) sizing rate. MDCTA correlated perfectly with DSA in the detection of intracranial aneurysms of anterior communicating artery and middle cerebral artery; however, slightly lower performance was noted in the PcoA and ICA supraclinoid segment aneurysms.


Vacuum-assisted closure of necrotic and infected cranial wound with loss of dura mater: A technical note.
Complex cranial wounds can be a problematic occurrence for surgeons. Vacuum-assisted closure devices have a wide variety of applications and have recently been used in neurosurgical cases involving complex cranial wounds. There is only one report regarding the use of a vacuum-assisted closure device with loss of dura mater. We report a complicated case of a necrotic cranial wound with loss of dura mater. A 68-year-old female underwent an evacuation of a subdural hematoma. Postoperatively, the patient developed a wound infection that required removal of the bone flap. The wound developed a wedge-shaped necrosis of the scalp with exposure of brain tissue due to loss of dura mater from previous surgeries. She underwent debridement and excision of the necrotic tissue with placement of a synthetic dural graft (Durepair®, Medtronic, Inc.) and placement of a wound vac. The patient underwent a latissismus dorsi muscle flap reconstruction that subsequently failed. After the wound vac was replaced, the synthetic dural graft was replaced with a fascia lata graft and an anterolateral thigh free flap reconstruction. We describe the technical nuances of this complicated case, how the obstacles were handled, and the literature that discusses the utility. We describe a case of a complex cranial wound and technical nuances on how to utilize a wound-vac with loss of dura mater.


A subcortical inhibitory signal for behavioral arrest in the thalamus.
Organization of behavior requires rapid coordination of brainstem and forebrain activity. The exact mechanisms of effective communication between these regions are presently unclear. The intralaminar thalamic nuclei (IL) probably serves as a central hub in this circuit by connecting the critical brainstem and forebrain areas. We found that GABAergic and glycinergic fibers ascending from the pontine reticular formation (PRF) of the brainstem evoked fast and reliable inhibition in the IL via large, multisynaptic terminals. This inhibition was fine-tuned through heterogeneous GABAergic and glycinergic receptor ratios expressed at individual synapses. Optogenetic activation of PRF axons in the IL of freely moving mice led to behavioral arrest and transient interruption of awake cortical activity. An afferent system with comparable morphological features was also found in the human IL. These data reveal an evolutionarily conserved ascending system that gates forebrain activity through fast and powerful synaptic inhibition of the IL.


Conformationally constrained peptides for drug delivery.
Peptides have shown great potential in acting as template for developing versatile carrier platforms in nanomedicine, aimed at selective delivery of drugs to only pathological tissues saving its normal neighbors. Cell-penetrating peptides (CPPs) are short oligomeric peptides capable of translocating across the cell membrane while simultaneously employing multiple mechanisms of entry. Most CPPs exist as disordered structures in solution and may adopt a helical conformation on interaction with cell membrane, vital to their penetrative capability. Herein, we report a series of cationic helical amphipathic peptides (CHAPs), which are topologically constrained to be helical. The peptides were tested against cervical and breast cancer cells for their cell penetration and drug delivery potential. The cellular uptake of CHAP peptides is independent of temperature and energy availability. The activity of the peptides is biocompatible in bovine serum. CHAPs delivered functional methotrexate (MTX) inside the cell as CHAP-MTX conjugates. CHAP-MTX conjugates were more toxic to cancer cells than MTX alone. However, the CHAP-MTX conjugates were less toxic to HEK-293 cells compared with the cancer cells suggesting higher affinity towards cancer cells.


Layer 4 of mouse neocortex differs in cell types and circuit organization between sensory areas.
Layer 4 (L4) of mammalian neocortex plays a crucial role in cortical information processing, yet a complete census of its cell types and connectivity remains elusive. Using whole-cell recordings with morphological recovery, we identified one major excitatory and seven inhibitory types of neurons in L4 of adult mouse visual cortex (V1). Nearly all excitatory neurons were pyramidal and all somatostatin-positive (SOM+) non-fast-spiking interneurons were Martinotti cells. In contrast, in somatosensory cortex (S1), excitatory neurons were mostly stellate and SOM+ interneurons were non-Martinotti. These morphologically distinct SOM+ interneurons corresponded to different transcriptomic cell types and were differentially integrated into the local circuit with only S1 neurons receiving local excitatory input. We propose that cell type specific circuit motifs, such as the Martinotti/pyramidal and non-Martinotti/stellate pairs, are used across the cortex as building blocks to assemble cortical circuits.


Naloxonazine antagonism of levorphanol-induced antinociception and respiratory depression in rhesus monkeys.
The mu-opioid receptor antagonist effects of naloxonazine on levorphanol-induced thermal antinociception and respiratory depression were examined in rhesus monkeys. Levorphanol (0.032-3.2 mg/kg) produced dose-dependent increases in tail-withdrawal latencies from 50 degrees C water in a warm-water tail-withdrawal assay and dose-dependent decreases in ventilation in both air and 5% CO2 mixed in air. Naloxonazine (0.1-3.0 mg/kg) antagonized both the antinociceptive and ventilatory effects of levorphanol to a similar degree, and the antagonist effects of naloxonazine were greater after 1 h than after 24 h. Under all conditions, the antagonist effects of naloxonazine were fully surmountable. Schild analysis of the antagonist effects of naloxonazine after 1 h pretreatment in the antinociception assay yielded a pA2 value of 7.6 and a slope of -0.50; by comparison, quadazocine yielded a pA2 value of 7.5 and a slope of -1.05. These results suggest that naloxonazine acts as a potent and fully reversible mu-opioid receptor antagonist with a moderately long duration of action in rhesus monkeys. In addition, these results suggest that the antinociceptive and ventilatory effects of mu-opioid receptor agonists in rhesus monkeys are mediated by pharmacologically similar populations of mu opioid receptors.


Oncologists' Selection of Genetic and Molecular Testing in the Evolving Landscape of Stage II Colorectal Cancer.
Little is known about the roles of genetic and molecular testing and Lynch syndrome screening in the formulation of predictive and prognostic assessments for patients with stage II colorectal cancer (CRC). From 2012 to 2013, we surveyed medical oncologists in the Cancer Care Outcomes Research and Surveillance Consortium and evaluated oncologists' selection of microsatellite instability (MSI) and/or immunohistochemistry (IHC) for mismatch repair (MMR) proteins, germline testing for MMR genes, BRAF and KRAS mutation analysis, and Oncotype DX in stage II CRC. Physicians were randomly assigned to receive one of three vignettes that varied by strength of CRC family history. We used multivariable logistic regression to identify physician and practice characteristics associated with test selection. Among 327 oncologists, MSI and/or IHC for MMR proteins were most frequently selected (n = 205; 64%), with 82% versus 53% choosing MSI/IHC testing in patients with strong versus no CRC family history, respectively (adjusted odds ratio [OR], 3.87; 95% CI, 2.07 to 7.22). KRAS and Oncotype DX testing were chosen by 24% and 38% of oncologists, respectively. Graduates of non-US and Canadian medical schools and physicians compensated by fee-for-service or on the basis of productivity were more likely to choose KRAS testing versus those receiving salaries not on the basis of productivity (OR, 2.16; 95% CI, 1.17 to 3.99; and OR, 1.94; 95% CI, 1.02 to 3.66, respectively). Fee-for-service or productivity-based salaries were also associated with increased odds of Oncotype DX testing (OR, 2.04; 95% CI, 1.17 to 3.55). Among surveyed oncologists, we found undertesting and overtesting related to genetic and molecular testing and Lynch syndrome screening for patients with stage II CRC,highlighting the need for improved implementation, targeted education, and evaluation of organizational and financial arrangements to promote the appropriate use of such tests.


Predictable changes in left ventricular mass and function during ten years after valve replacement for aortic stenosis.
Left ventricular (LV) hypertrophy is the underlying basis for longevity after aortic valve replacement (AVR) for aortic stenosis (AS). However, a detailed account of changes in LV mass and function in the long term after AVR and identification of the determinants of such changes have not yet been presented. Ninety-one unselected consecutive adult patients with AS underwent AVR and were followed up to 10 years, at which time 41 survivors without new mitral disease underwent repeat measurement of LV mass index (LVMi), ejection fraction (LVEF), fast filling fraction (LVFFF), and end-diastolic volume index (LVEDVi). A subgroup comprising 49 patients was also assessed at eight days, three months, and 1.5 years postoperatively. All measurements were analyzed in a longitudinal regression model for repeated measures. LVMi fell from 202 +/- 58 g/m2 (n = 91) via 150 +/- 45 g/m2 (n = 39) at 1.5 years to 139 +/- 40 g/m2 (n = 41) at 10 years in all patients, and to 124 +/- 31 g/m2 (n = 29) in non-hypertensive patients. The LVMi falls were paralleled by improvements in LVEF and LVEDVi. LVFFF was not correlated to LVMi before the 10-year study. The longitudinal model indicated progressive reduction of LVMi to 1.5 years, but no change thereafter. The predictor variables were preoperative LVMi and end-systolic dimension index (high values of both related to high postoperative LVMi), hypertension, and male gender. The model for LVEF indicated a rapid increase to three months, followed by a slight decrease to 1.5 years and further to 10 years, predicted by preoperative LVEF and LVFFF. LVFFF fell sharply by three months, had recovered somewhat at 1.5 years and fully at 10 years, positively related to preoperative LVFFF and inversely to end-systolic chamber radius:wall thickness ration and small-sized prosthetic valves. LVEDVi converged from extreme values over time predicted by preoperative LVEF, but rose with hypertension and coronary artery disease. Hemodynamic function of the prosthetic aortic valve at any of the measurement times had no impact. Changes in LV mass and function up to 10 years after AVR for AS were highly predictable. Poorer outcomes were related to preoperative excessive hypertrophy and indices of underlying irreversible myocardial disease and further compromised by hypertension and, to a lesser extent, coronary artery disease. The hemodynamic function of the aortic prosthetic valve did not seem to play a role.


Interaction of organic chemicals with P-glycoprotein in the adrenal gland, kidney, and a multidrug-resistant KB cell.
P-glycoprotein (P-gp) is thought to mediate the transport of anti-cancer drugs and to be responsible for the multidrug-resistant (MDR) phenotype in tumor cells. However, the function of P-gp in normal tissues is still not well understood. We present evidence indicating that the active efflux of several structurally unrelated organic compounds is mediated by P-gp in multidrug-resistant KB (KB-C2) cells and that these compounds interact with P-gp in the kidney and adrenal gland. The photoactive radioactive calcium channel blocker [3H]azidopine labels a protein of approximately 140 kDa in crude membrane fractions from human kidney and adrenal gland and a 130-kDa protein from bovine adrenal gland. These photolabeled proteins are immunoprecipitated with an anti-P-gp antibody. Photolabeling is inhibited by vinblastine, reserpine, and several other organic chemicals. These data indicate that the kidney and adrenal gland express P-gp (or a protein closely related to P-gp) that can interact with several organic compounds and that the P-gp expressed in these tissues has a drug-binding site similar to that of P-gp in KB-C2 cells. Our findings thus strongly support the hypothesis that P-gp can transport a wide variety of organic chemicals as well as anti-cancer drugs and that one of the physiological functions of P-gp is the excretion of certain classes of organic compounds.


Hemangioblastoma of the cervicomedullary junction. Report of three cases.
Solid hemangioblastomas of the cervicomedullary junction are benign vascular neoplasms malignantly located. Their unusual and characteristic clinical presentations include loss of involuntary respiratory drive, neurogenic hypertension, and emesis; these are secondary to their involvement of the tractus solitarius in the medulla. Although angiography is diagnostic, the computerized tomography appearance and unusual location also suggest the diagnosis. The correct preoperative diagnosis is particularly important for the successful surgical excision of these vascular tumors. The microsurgical removal of cervicomedullary hemangioblastomas is described in three patients.


Multiple effects induced by unstable mutation in Streptomyces lavendulae.
Arg mutants, isolated from Streptomyces lavendulae at unusually high frequencies, showed several phenotypic characteristics. The characteristics common to all arg mutants include: (1) repression of beta-lactamase production, (2) inhibition of aerial mycelium formation, (3) development of acid pH, (4) low saturation density of growth in liquid culture, (5) a decrease in antibiotic production, (6) an increase in sensitivity to benzylpenicillin and (7) a decrease in production of pigment. These results suggest that the arg mutation concomitantly caused the depression of secondary metabolism in S. lavendulae.


'Primer alignment-and-extension': a novel mechanism of viral RNA recombination responsible for the rescue of inactivated poliovirus cDNA clones.
In the course of experiments designed to assess the potential role of alternative open reading frames (ORF) present in the 5'-terminal untranslated region (5'-UTR) of poliovirus type 1 (Mahoney strain) genomic RNA, we came across a double mutation that completely abrogated the infectivity of full-length cDNA clones. The infectivity was rescued in trans by cotransfecting COS-1 cells with short RNA transcripts of the wild-type 5'-UTR of poliovirus type 2 Lansing, provided a free 3'-OH was available. Direct sequencing of the viral RNA revealed that the infectious viruses recovered were recombinants Lansing/Mahoney, with variable points of 'crossing-over'. A novel mechanism of RNA-RNA recombination, which we propose to call 'primer alignment-and-extension', is described that would explain the high rate of recombination of RNA viruses observed in natural conditions.


[The homeostasis assessment by hemorrhagic shock].
The comparative analysis of different protocols of infusion therapy of the hemorrhagic shock stage III was performed. The infusion of the colloid solution of hydroxiethylstarch 200/0.5 and non-balanced crystalloid 0.9% solution of natrium chloride leads to the development of negative changes in homeostasis. Whereas infusion of the 4% solution of the modified gelatin and balanced crystalloid solition (sterofundin) allows to avoid the registered changes in electrolyte and alkaline balance.


Probing molecular frame photoionization via laser generated high-order harmonics from aligned molecules.
Present experiments cannot measure molecular frame photoelectron angular distributions (MFPAD) for ionization from the outermost valence orbitals of molecules. We show that the details of MFPAD can be retrieved with high-order harmonics generated by infrared lasers from aligned molecules. Using accurately calculated photoionization transition dipole moments for fixed-in-space molecules, we show that the dependence of the magnitude and phase of the high-order harmonics on the alignment angle of the molecules observed in recent experiments can be quantitatively reproduced. This result provides the needed theoretical basis for ultrafast dynamic chemical imaging using infrared laser pulses.


Free tissue transfer to the lower extremity: a paradigm shift in flap selection for soft tissue reconstruction.
The free anterolateral thigh (ALT) skin perforator flap has gained enormous popularity as a free skin perforator flap. Currently being used for various reconstructions primarily in Asia, its routine application for soft-tissue reconstruction of the lower extremity remains unknown in the United States. The purpose of the present study was to review the current trend of flap selection for soft tissue reconstruction of the lower extremity. We performed a retrospective review of a single-surgeon case log between 2007 and 2011 in a tertiary academic medical center. Inclusion criteria were patients who underwent free tissue transfer for lower extremity reconstruction regardless of sex or age. Exclusion criteria were limited to patients with a body mass index greater than 35 kg/m. A total of 20 patients underwent reconstruction with a free tissue transfer for soft tissue coverage of the lower extremity wound were identified. Fourteen free ALT and 6 free muscle flap reconstructions were performed over a 4-year period. Good reconstructive outcomes resulting in reliable soft tissue coverage were achieved in all patients. It was noted that more ALT flaps were used in the last 2 years and less muscle flaps (0/8 vs 6/12, P = 0.042) when compared with the years prior. Although technically more challenging to perform, the free ALT flap has recently been used significantly more for lower extremity reconstruction because of its minimal donor-site morbidity, reliable coverage, and improved contour. This represents a paradigm shift in flap selection that is being highlighted as surgical proficiency in flap dissection technique improves.


Population-Sequencing as a Biomarker of Burkholderia mallei and Burkholderia pseudomallei Evolution through Microbial Forensic Analysis.
Large-scale genomics projects are identifying biomarkers to detect human disease. B. pseudomallei and B. mallei are two closely related select agents that cause melioidosis and glanders. Accurate characterization of metagenomic samples is dependent on accurate measurements of genetic variation between isolates with resolution down to strain level. Often single biomarker sensitivity is augmented by use of multiple or panels of biomarkers. In parallel with single biomarker validation, advances in DNA sequencing enable analysis of entire genomes in a single run: population-sequencing. Potentially, direct sequencing could be used to analyze an entire genome to serve as the biomarker for genome identification. However, genome variation and population diversity complicate use of direct sequencing, as well as differences caused by sample preparation protocols including sequencing artifacts and mistakes. As part of a Department of Homeland Security program in bacterial forensics, we examined how to implement whole genome sequencing (WGS) analysis as a judicially defensible forensic method for attributing microbial sample relatedness; and also to determine the strengths and limitations of whole genome sequence analysis in a forensics context. Herein, we demonstrate use of sequencing to provide genetic characterization of populations: direct sequencing of populations.


Epitaxy: Programmable Atom Equivalents Versus Atoms.
The programmability of DNA makes it an attractive structure-directing ligand for the assembly of nanoparticle (NP) superlattices in a manner that mimics many aspects of atomic crystallization. However, the synthesis of multilayer single crystals of defined size remains a challenge. Though previous studies considered lattice mismatch as the major limiting factor for multilayer assembly, thin film growth depends on many interlinked variables. Here, a more comprehensive approach is taken to study fundamental elements, such as the growth temperature and the thermodynamics of interfacial energetics, to achieve epitaxial growth of NP thin films. Both surface morphology and internal thin film structure are examined to provide an understanding of particle attachment and reorganization during growth. Under equilibrium conditions, single crystalline, multilayer thin films can be synthesized over 500 × 500 μm2 areas on lithographically patterned templates, whereas deposition under kinetic conditions leads to the rapid growth of glassy films. Importantly, these superlattices follow the same patterns of crystal growth demonstrated in atomic thin film deposition, allowing these processes to be understood in the context of well-studied atomic epitaxy and enabling a nanoscale model to study fundamental crystallization processes. Through understanding the role of epitaxy as a driving force for NP assembly, we are able to realize 3D architectures of arbitrary domain geometry and size.


Identifying the subwavelength-aperture-width variation in the far field with tunable asymmetrically-embedded-aperture interferometer configuration.
A tunable asymmetrically-embedded-aperture interferometer configuration is proposed to enhance the measurement sensitivity of subwavelength variation. With this configuration, an aperture of reference was posited asymmetrically, relative to the test aperture, that exhibited subwavelength variation. By a shift in the relative position of the reference aperture, the detection sensitivity of measuring the subwavelength variation in the far field can be enhanced to a desired value at any specific detection width. We discuss the underlying mechanism of optimization and address the difference between the embedded-aperture interferometry and tunable asymmetrically-embedded-aperture interferometry with respect to tolerance.


Age-dependent plasticity in endocannabinoid modulation of pain processing through postnatal development.
Significant age- and experience-dependent remodelling of spinal and supraspinal neural networks occur, resulting in altered pain responses in early life. In adults, endogenous opioid peptide and endocannabinoid (ECs) pain control systems exist which modify pain responses, but the role they play in acute responses to pain and postnatal neurodevelopment is unknown. Here, we have studied the changing role of the ECs in the brainstem nuclei essential for the control of nociception from birth to adulthood in both rats and humans. Using in vivo electrophysiology, we show that substantial functional changes occur in the effect of microinjection of ECs receptor agonists and antagonists in the periaqueductal grey (PAG) and rostroventral medulla (RVM), both of which play central roles in the supraspinal control of pain and the maintenance of chronic pain states in adulthood. We show that in immature PAG and RVM, the orphan receptor, GPR55, is able to mediate profound analgesia which is absent in adults. We show that tissue levels of endocannabinoid neurotransmitters, anandamide and 2-arachidonoylglycerol, within the PAG and RVM are developmentally regulated (using mass spectrometry). The expression patterns and levels of ECs enzymes and receptors were assessed using quantitative PCR and immunohistochemistry. In human brainstem, we show age-related alterations in the expression of key enzymes and receptors involved in ECs function using PCR and in situ hybridisation. These data reveal that significant changes on ECs that to this point have been unknown and which shed new light into the complex neurochemical changes that permit normal, mature responses to pain.


Mycobacterium tuberculosis Infection in Close Childhood Contacts of Adults with Pulmonary Tuberculosis is Increased by Secondhand Exposure to Tobacco.
Tobacco use is a major risk factor for tuberculosis (TB). Secondhand smoke (SHS) is also a risk factor for TB and to a lesser extent, Mycobacterium tuberculosis infection without disease. We investigated the added risk of M. tuberculosis infection due to SHS exposure in childhood contacts of TB cases in The Gambia. Participants were childhood household contacts aged ≤ 14 years of newly diagnosed pulmonary TB (PTB) cases. The intensity of exposure to the case was categorized according to whether contacts slept in the same room, same house, or a different house as the case. Contacts were tested with an enzyme-linked immunospot interferon gamma release assay. In multivariate regression models, M. tuberculosis infection was associated with increasing exposure to a case (odds ratios [OR]: 3.9, 95% confidence interval [CI]: 2.11-71.4, P < 0.001]) and with male gender (OR: 1.5 [95% CI: 1.12-2.11], P = 0.008). Tobacco use caused a 3-fold increase in the odds of M. tuberculosis infection in children who slept closest to a case who smoked within the same home compared with a nonsmoking case (OR: 8.0 [95% CI: 2.74-23.29] versus 2.4 [95% CI: 1.17-4.92], P < 0.001). SHS exposure as an effect modifier appears to greatly increase the risk of M. tuberculosis infection in children exposed to PTB cases. Smoking cessation campaigns may be important for reducing transmission of M. tuberculosis to children within households.


A case of resistance to tyrosine kinase inhibitor therapy: small cell carcinoma transformation concomitant with plasma-genotyped T790M positivity.
Although non-small-cell lung cancer patients with epidermal growth factor receptor (EGFR) mutations are responsive to EGFR-tyrosine kinase inhibitors, drug resistances are always inevitable. The secondary somatic EGFR threonine-methionine substitution at position 790 (T790M) mutation accounts for ∼50% of acquired resistance mechanisms. Small cell lung cancer (SCLC) transformation is a relatively rare mechanism, but has recently attracted considerable attention. The coexistence of both the mechanisms in one patient is much more scarce in clinic. In this case report, we described a 37-year-old woman who underwent refractory after second-line gefitinib therapy and was confirmed to have SCLC transformation without the T790M mutation in the left lobar nodule, but concomitant with the plasma-genotyped EGFR T790M mutation. Our case report uncovered the underling relationship between SCLC transformation and the T790M mutation, and the fluid biopsy approach may help overcome the problem of heterogeneity in acquired resistance to EGFR-tyrosine kinase inhibitors.


Elmer's glue, Elsie and you: clinical applications of adhesion molecules.
An effective response to injury or inflammation requires leukocyte migration from the endovascular compartment into areas of inflammation. This process requires the appropriate expression of adhesion molecules, which mediate cell-cell and cell-extracellular matrix interactions. A better understanding of the roles of adhesion molecules in normal and pathological conditions might lead to the development of more effective therapeutic interventions. Features of several classes of adhesion molecules, including immunoglobulin superfamily, selectins, and integrins, are reviewed along with how they participate in leukocyte-endothelial interactions. The role of cell adhesion molecules in a variety of pathological conditions, including graft rejection, reperfusion injury, and acute lung injury are discussed. In addition, some recent studies that explore therapeutic uses of adhesion molecules are summarized.


A sensitive and selective molecularly imprinted sensor combined with magnetic molecularly imprinted solid phase extraction for determination of dibutyl phthalate.
A highly sensitive and selective molecularly imprinted (MIP) sensor combined with magnetic molecularly imprinted solid phase extraction (MMISPE) was developed for the determination of dibutyl phthalate (DBP) in complex matrixes. The magnetic molecularly imprinted polymer (MMIP) was synthesized as solid phase extraction (SPE) sorbet to extract DBP from complex matrixes and as sensing element to improve the selectivity of the imprinted sensor. The morphologies of MMIP and MIP-sensor were characterized by using scanning electron microscope (SEM) and transmission electron microscopy (TEM). The electrochemical performances of MIP-sensor were investigated by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The conditions of preconcentration, elution and electrochemical determination were studied in detail. Under the optimized experimental conditions, the response currents of the MIP-sensor exhibited a linear relationship towards DBP concentrations ranging from 1.0 × 10(-8)g/L to 1.0 × 10(-3)g/L. The limit of detection of the MMIP-sensor coupled with the MMISPE was calculated as 0.052 ng/L. The MMIP-sensor coupled with the MMISPE was applied to detect DBP in complex samples successfully.


Correlation of serum tryptase levels with total number of nevi, Breslow thickness, ulceration, and mitotic index in melanoma patients: evaluation of a promising prognostic marker.
Current evidences suggest that mast cells contribute to the proliferation and differentiation of skin melanocytes. According to these findings, we carried out an observational cross-sectional study to investigate the correlation between the total number of nevi (TN), Breslow thickness (BT), and serum tryptase (ST) levels in a cohort of 35 melanoma (MM) patients. A Mann-Whitney test was performed to compare ST values within each variable. Subsequently, the independent predictive factors were assessed by multiple logistic regression. Pearson's χ-test was chosen to detect statistically significant findings on the TN and the histopatological variables (Breslow, ulceration, and mitotic index). The TN was assessed using a dichotomous scale (≤ 10 or > 10). Patients with TN of 10 or less (3.48 vs. 6.05 ng/ml; P = 0.045), patients with a Breslow thickness of at least 1.01 mm (2.99 vs. 5.67 ng/ml; P = 0.1), and ulcerated MM (2.37 vs. 6.05 ng/ml; P < 0.001) showed lower median ST levels. Similarly, MM with mitotic index of at least 1/mm had median ST levels lower than MM with mitotic index less than 1/mm (P = 0.005). Multiple logistic regression confirmed the statistical significance for the variables ulceration, TN, and mitotic index. Pearson's χ-test showed a statistically significantly (P = 0.003) increased prevalence of MMs with a BT of at least 1.01 mm in patients with a TN of 10 or less. Patients with a TN of 10 or less also showed a higher prevalence of ulceration and mitotic index of at least 1/mm in comparison with the rest of the cohort. Our study highlights lower median ST levels in patients whose MM thickness is at least 1.01 mm; this may encourage new studies on the role of ST in MM also according to the number of nevi.


Lethal toxoplasmosis in an aviary of kakarikis (Cyanoramphus spp.) in Australia.
A small outbreak of fatal toxoplasmosis is described in 6 kakariki (Cyanoramphus spp.) in an aviary in Australia. Toxoplasma gondii-like tachyzoites were found associated with necrosis in intestine, spleen, liver, and lungs. Protozoal tachyzoites reacted positively with T. gondii-specific polyclonal antibodies. This is the first report of toxoplasmosis in kakariki.


Leisure-time physical activity over the life course and cognitive functioning in late mid-adult years: a cohort-based investigation.
The objective of the present study was to estimate the association between different leisure-time physical activity (LTPA) parameters from 11 to 50 years and cognitive functioning in late mid-adulthood. The study used a prospective birth cohort study including participants in the UK National Child Development Study (NCDS) from age 11 to 50 years. Standardized z scores for cognitive, memory and executive functioning at age 50 represented the primary outcome measures. Exposures included self-reported LTPA at ages 11, 16, 33, 42, 46 and 50 years. Analyses were adjusted for important confounders including educational attainment and long-standing illness. The adjusted difference in cognition score between women who reported LTPA for at least 4 days/week in five surveys or more and those who never reported LTPA for at least 4 days/week was 0.28 [95% confidence interval (CI) 0.20-0.35], 0.10 (95% CI 0.01-0.19) for memory score and 0.30 (95% CI 0.23-0.38) for executive functioning score. For men, the equivalent differences were: cognition 0.12 (95% CI 0.05-0.18), memory 0.06 (95% CI -0.02 to 0.14) and executive functioning 0.16 (95% CI 0.10-0.23). This study provides novel evidence about the lifelong association between LTPA and memory and executive functioning in mid-adult years. Participation in low-frequency and low-intensity LTPA was positively associated with cognitive functioning in late mid-adult years for men and women. The greatest benefit emerged from participating in lifelong intensive LTPA.


Pathogenesis of pancreatic and biliary CaCO3 lithiasis: the solubility product (K'sp) of calcite determined with the Ca++ electrode.
There are three digestive secretory fluids with high bicarbonate and high pH values: bile, pancreatic juice, and saliva. Each is subject to development of CaCO3-containing stones. In bile, calcium precipitation is a requisite event in the initiation and growth of all pigment gallstones and is postulated to initiate cholesterol gallstone formation by forming a nidus for cholesterol precipitation. In the pancreas, stones in both humans and cattle are composed largely of calcite. Knowledge of appropriate solubility product constant (K'sp) values is essential in defining lithogenicity in each of these secretions. Only two studies of calcite solubility at physiologic total ionic strength (u) have been found, both of which used total calcium measurements to estimate free Ca++ ion in calculating K'sp. The Ca++ electrode has allowed a fresh appraisal of this problem. Studies of calcite were made at 24 degrees C and 37 degrees C and u = 0.16 mol/L over an 11-day period. At each temperature, pH declined slightly with time, reflecting atmospheric CO2 uptake. As predicted by theory, this decline was associated with increase in free Ca++ ion and total calcium concentration, but K'sp remained constant because of corresponding decline in [CO=3]. Mean K'sp at 24 degrees C was 1.33 +/- 0.04 X 10(-8) mol/L, and at 37 degrees C it was 3.76 +/- 0.09 X 10(-8) mol/L. A lithogenicity diagram, applicable to bile, pancreatic juice, saliva, plasma, and other body fluids at u = 0.16 mol/L, is presented for [Ca++]sat, the free Ca++ ion concentration at the limit of a stable thermodynamic state.(ABSTRACT TRUNCATED AT 250 WORDS)


Screening of medicinal plant phytochemicals as natural antagonists of p53-MDM2 interaction to reactivate p53 functioning.
In most types of cancer, overexpression of murine double minute 2 (MDM2) often leads to inactivation of p53. The crystal structure of MDM2, with a 109-residue amino-terminal domain, reveals that MDM2 has a core hydrophobic region to which p53 binds as an amphipathic α helix. The interface depends on the steric complementarity between MDM2 and the hydrophobic region of p53. Especially, on p53's triad, amino acids Phe19, Trp23 and Leu26 bind to the MDM2 core. Results from studies suggest that the structural motif of both p53 and MDM2 can be attributed to similarities in the amphipathic α helix. Thus, in the current investigation it is hypothesized that the similarity in the structural motif might be the cause of p53 inactivation by MDM2. Hence, molecular docking and phytochemical screening approaches are appraised to inhibit the hydrophobic cleft of MDM2 and to stop p53-MDM2 interaction, resulting in reactivation of p53 activity. For this purpose, a library of 2295 phytochemicals were screened against p53-MDM2 to find potential candidates. Of these, four phytochemicals including epigallocatechin gallate, alvaradoin M, alvaradoin E and nordihydroguaiaretic acid were found to be potential inhibitors of p53-MDM2 interaction. The screened phytochemicals, derived from natural extracts, may have negligible side effects and can be explored as potent antagonists of p53-MDM2 interactions, resulting in reactivation of the normal transcription of p53.


Economic credentialing is here to stay.
Health care organizations are being scrutinized by payers for the efficiency of their processes to render health care. Organizations must offer alternative avenues to satisfy health care needs that are less resource consuming and have a reasonable chance of success. This presents an enormous challenge to U.S. health care. In the past, while in training, physicians were conditioned to ignore costs in the provision of care. We cannot afford that behavior today. Physicians must be reeducated and their behavior reconditioned to alter the teachings at medical school and residency with respect to resource utilization. To be effective, this education and behavior modification must be done in a nonpunitive fashion.


Ribosomal DNA analysis of tsetse and non-tsetse transmitted Ethiopian Trypanosoma vivax strains in view of improved molecular diagnosis.
Animal trypanosomosis caused by Trypanosoma vivax (T. vivax) is a devastating disease causing serious economic losses. Most molecular diagnostics for T. vivax infection target the ribosomal DNA locus (rDNA) but are challenged by the heterogeneity among T. vivax strains. In this study, we investigated the rDNA heterogeneity of Ethiopian T. vivax strains in relation to their presence in tsetse-infested and tsetse-free areas and its effect on molecular diagnosis. We sequenced the rDNA loci of six Ethiopian (three from tsetse-infested and three from tsetse-free areas) and one Nigerian T. vivax strain. We analysed the obtained sequences in silico for primer-mismatches of some commonly used diagnostic PCR assays and for GC content. With these data, we selected some rDNA diagnostic PCR assays for evaluation of their diagnostic accuracy. Furthermore we constructed two phylogenetic networks based on sequences within the smaller subunit (SSU) of 18S and within the 5.8S and internal transcribed spacer 2 (ITS2) to assess the relatedness of Ethiopian T. vivax strains to strains from other African countries and from South America. In silico analysis of the rDNA sequence showed important mismatches of some published diagnostic PCR primers and high GC content of T. vivax rDNA. The evaluation of selected diagnostic PCR assays with specimens from cattle under natural T. vivax challenge showed that this high GC content interferes with the diagnostic accuracy of PCR, especially in cases of mixed infections with T. congolense. Adding betain to the PCR reaction mixture can enhance the amplification of T. vivax rDNA but decreases the sensitivity for T. congolense and Trypanozoon. The networks illustrated that Ethiopian T. vivax strains are considerably heterogeneous and two strains (one from tsetse-infested and one from tsetse-free area) are more related to the West African and South American strains than to the East African strains. The rDNA locus sequence of six Ethiopian T. vivax strains showed important differences and higher GC content compared to other animal trypanosomes but could not be related to their origin from tsetse-infested or tsetse-free area. The high GC content of T. vivax DNA renders accurate diagnosis of all pathogenic animal trypanosomes with one single PCR problematic.


Effect of tif expression, irradiation of recipient and presence of plasmid pKM101 on recovery of a marker from a donor exposed to ultraviolet light prior to conjugation.
To detect the effect of the postulated inducible error-prone repair system ('SOS repair') on the bacterial chromosome, an Hfr Escherichia coli strain JC5088 recA was u.v.-irradiated immediately before mating it with recipients in which SOS repair was supposed to be functioning through tif expression, u.v. irradiation or the presence of plasmid pKM101. The recombinant yields of these crosses were compared with those obtained in corresponding crosses with recipients in which SOS repair either was not induced or was totally eliminated by the lexA mutation. No difference in marker recovery efficiency could be detected between these two sets of recipients and thus no induced repair process acting on donor DNA could be demonstrated. The possible reasons for this finding are discussed.


Do local enhanced services in primary care improve outcomes? Results from a literature review.
This paper aims to examine the role of local enhanced services (LES) as a financial incentive in improving clinical and process outcomes in primary care with a view to discussing their future in light of the Health and Social Care Act. A literature review was conducted to identify LES commissioned in the UK in any disease area and to evaluate common themes relating to their impact on outcomes. The literature review consisted of two stages: an initial reference database search (MEDLINE, MEDLINE IN-PROCESS and EMBASE) and a more general internet search. The internet search used free text augmented by a targeted search of key health organisations' websites. Data were extracted from the LES to provide information on the background and context of the LES before going on to describe the incentive structure, health and economic outcomes and limitations of the LES. Although a number of LES were identified in the online search, only 14 reported any data on outcomes. These LES programmes related to 10 different disease areas, with cancer, alcohol dependence and chronic obstructive pulmonary disease (COPD) being the most common health needs. Three common factors between the selected LES emerged that appear to influence the extent of the impact on local health or economic outcomes: (1) a national framework supporting the LES, (2) existing service provision, and (3) the size of the financial incentives. The common themes emerging from the literature review suggest that, following the Health and Social Care Act 2012 and newly established national standards, given sufficient attention to planning service specifications, LES could continue to be important in reducing health inequalities and preparing poorly performing general practices for longer term changes directed at improving outcomes and standards in healthcare.


Effects of oxatomide compared with chlorpheniramine in allergic rhinoconjunctivitis.
Oxatomide is an H1 receptor antagonist, which also inhibits antigen-induced mediator release from sensitized mast cells and basophils. The effects of ingested oxatomide (30 mg/capsule) were compared with those of chlorpheniramine (5 mg/capsule) in a double-blind non-crossover study in two groups of twenty-five subjects with ragweed pollen-induced hay fever during the pollen season. Oxatomide or chlorpheniramine was started when symptoms became troublesome and was used in a dose of two to four capsules daily according to severity. Other medications were added, in a standardized way, if hay fever symptoms were not controlled by the test capsules. Forty-one subjects completed the study; nineteen received oxatomide and twenty-two chlorpheniramine. Mean symptom scores, number of test capsules and number of doses of additional medication were not significantly different between the two groups. Drowsiness was reported by subjects in both groups but was worse in the chlorpheniramine group (P less than 0.05). We conclude that, in the studied doses, oxatomide is as effective for the treatment of allergic rhinoconjunctivitis as chlorpheniramine and produces less drowsiness.


A Pseudo-Outbreak of Pseudomonas putida and Stenotrophomonas maltophilia in a Bronchoscopy Unit.
Endoscopes represent the medical devices most commonly linked to health care-associated outbreaks and pseudo-outbreaks. Most of the recent outbreaks and pseudo-outbreaks have resulted from contaminated automated endoscope reprocessors (AER) or the use of damaged or malfunctioning bronchoscopes or contaminated equipment. We investigated a pseudo-outbreak of Pseudomonas putida and Stenotrophomonas maltophilia recovered from bronchial washing (BW) specimens obtained during bronchoscopy in a bronchoscopy unit. Samples were obtained from environmental surfaces in the endoscopy suite, bronchoscopes, and bronchoscopic dispensable material, and specimens of cleaning solutions, cleaning brushes, the AER, and the ultrasound system were sent for bacterial culture. Medical records were reviewed to identify possible infections after a bronchoscopy. P. putida and S. maltophilia were isolated from BW samples of 39 patients. The bronchoscopy models Olympus BF-1T160 and BF-160 were contaminated. Both bronchoscopes and other contaminated material (cleaning brushes, diluted cleaning solutions, and the sink) were isolated, but new cases continued to appear. The AER was recently installed, and new connections were used for the water lines and new tubes were connected to the AER. Initially, specimens were obtained from the external circuits and the internal walls of the AER. Finally, cultures were made from the filters on the water lines, and growth of P. putida and S. maltophilia was found. The investigation revealed that the BW specimens were contaminated because sterile saline was injected by means of the biopsy port of the bronchoscope and was recovered through the same channel by means of the proximal suction port. No patients developed clinical signs or symptoms of infection, but the positive cultures did lead to treatment of 21 patients. We described a pseudo-outbreak related to a contaminated bronchoscope because of inadequate installation of the AER for used new water lines and because the new tubes were connected to the AER. The antibacterial filters of the AER used tap water, and this may have contained low levels of microorganisms. No serious clinical complications derived from this pseudo-outbreak.


From cultivation to cancer: formation of N-nitrosamines and other carcinogens in smokeless tobacco and their mutagenic implications.
Tobacco use is a major cause of preventable morbidity and mortality globally. Tobacco products, including smokeless tobacco (ST), generally contain tobacco-specific N-nitrosamines (TSNAs), such as N'-nitrosonornicotine (NNN) and 4-(methylnitrosamino)-1-(3-pyridyl)-butanone (NNK), which are potent carcinogens that cause mutations in critical genes in human DNA. This review covers the series of biochemical and chemical transformations, related to TSNAs, leading from tobacco cultivation to cancer initiation. A key aim of this review is to provide a greater understanding of TSNAs: their precursors, the microbial and chemical mechanisms that contribute to their formation in ST, their mutagenicity leading to cancer due to ST use, and potential means of lowering TSNA levels in tobacco products. TSNAs are not present in harvested tobacco but can form due to nitrosating agents reacting with tobacco alkaloids present in tobacco during certain types of curing. TSNAs can also form during or following ST production when certain microorganisms perform nitrate metabolism, with dissimilatory nitrate reductases converting nitrate to nitrite that is then released into tobacco and reacts chemically with tobacco alkaloids. When ST usage occurs, TSNAs are absorbed and metabolized to reactive compounds that form DNA adducts leading to mutations in critical target genes, including the RAS oncogenes and the p53 tumor suppressor gene. DNA repair mechanisms remove most adducts induced by carcinogens, thus preventing many but not all mutations. Lastly, because TSNAs and other agents cause cancer, previously documented strategies for lowering their levels in ST products are discussed, including using tobacco with lower nornicotine levels, pasteurization and other means of eliminating microorganisms, omitting fermentation and fire-curing, refrigerating ST products, and including nitrite scavenging chemicals as ST ingredients.


Open trial of flutamide for treatment of obsessive-compulsive disorder.
Several lines of evidence suggest that gonadal steroid hormones play a role in the onset and exacerbation of obsessive-compulsive disorder (OCD). In this study, we examined the effects of treatment with flutamide, a synthetic, nonsteroidal, competitive antagonist of the androgen receptor, on OCD symptoms. Eight outpatients meeting DSM-III-R criteria for OCD participated in an 8-week open trial of flutamide. The dose was increased from 250 mg/day to 750 mg/day over the first 4 weeks and maintained at 750 mg/day for the final 4 weeks. The primary outcome measures for OCD symptoms were the Yale-Brown Obsessive Compulsive Scale and the Maudsley Inventory and for anxiety symptoms, the Beck Anxiety Inventory and the Hamilton Rating Scale for Anxiety. Subjects also provided self-ratings of aggression and sexual interest and activity. There were no reductions in measures of obsession and compulsions or measures of anxiety over the 8-week trial. However, self-ratings of feelings of aggression did fall significantly over the 8-week trial (p < .001). The lack of response to treatment with flutamide, an androgen receptor antagonist, suggests that any effects of gonadal steroids to exacerbate OCD symptoms are more likely to be mediated through estrogen receptors or through mechanisms that do not involve classical intracellular androgen receptors. Future treatment trials should examine agents that antagonize estrogen receptors or otherwise inhibit estrogen activity.


Nutrition and immunity: the influence of diet on autoimmunity and the role of zinc in the immune response.
Nutrition exerts profound influence on immunological functions effecting both cell-mediated (humoral) and T cell-mediated (cellular) immune functions. Even the interaction of the immune systems can be profoundly influenced by restrictions or excesses of dietary constituents. In experimental systems where it is possible to control precisely the influence of specific nutriments, development and expression of autoimmune diseases and the associated immunodeficiencies of aging can be delayed by restrictions of dietary protein, protein and calories, fat, zinc, or even essential fatty acids. Tumor immunities likewise can be affected and sometimes even enhanced by restriction of protein, calories, or protein and calories, an influence associated with major delay in development of the experimental cancers--e.g. breast cancer. T cell-mediated immunodeficiencies associated with clinically apparent protein or protein calorie malnutrition are often attributable not to the major nutriment deficiencies per se but to accompanying zinc deficiency, a finding reflecting the vital role of zinc in many immunological functions. Dietary zinc deficiency appears to be responsible, at least in part, for the immunodeficiency that is so regularly associated with certain human cancers, such as epidermoid cancers of the head and neck region.


Differential effects of macronutrient content in 2 energy-restricted diets on cardiovascular risk factors and adipose tissue cell size in moderately obese individuals: a randomized controlled trial.
The most effective and safe dietary approach for weight loss and its impact on the metabolic functions and morphology of adipose tissue remain unclear. We evaluated whether an energy-restricted high-protein diet with a low glycemic index and soluble fiber (LC-P-LGI) would be more effective than a low-calorie conventional diet (LC-CONV) on weight loss and related metabolic risk factors. We further determined factors that may influence adipocyte size during energy restriction. Thirteen obese participants were randomly assigned in a crossover design to 2 periods of a 4-wk hypocaloric diet as either LC-P-LGI or LC-CONV, separated by 8-wk washout intervals. In comparison with the LC-CONV diet, the main effect of the LC-P-LGI diet was a greater decrease in adipocyte diameter (P = 0.048), plasma plasminogen activator inhibitor protein-1 (P = 0.019), vascular endothelial growth factor (P = 0.032), and interferon-γ inducible protein 10 (P = 0.010). Whereas fasting plasma glucose and high-sensitivity C-reactive protein decreased only after the LC-P-LGI diet, with no differences between diets, fasting plasma insulin and insulin resistance were lower after the LC-CONV diet. The diet results did not differ for body composition and lipid variables. Kinetic modifications in adipocyte diameter were associated with metabolic variables and genes implicated in adipocyte proliferation, apoptosis, and angiogenesis. In comparison with the LC-CONV diet, the LC-P-LGI diet was associated with improvement in some cardiometabolic risk factors and greater reduction in adipocyte size. Profiles of genes involved in inhibiting adipogenesis and angiogenesis, but increasing apoptosis, were correlated with decreased adipocyte size. This study provides insight into the adipose tissue-remodeling changes that induce regulation of adipocyte size during dietary weight loss. This trial was registered at clinicaltrials.gov as NCT01312740.


Preventive approach to tobacco-related diseases.
The prevention of tobacco-related diseases should involve the combined efforts of many different segments of science, medicine, education, and of society as such. It needs to begin with efforts to prevent children from starting to smoke. School health education on smoking and other risk factors that relate to health, need to become an integral and consistent part of school education. We shall demonstrate from the KYB (Know Your Body) school health education program, how such efforts could be advanced. In respect to the adult population, media must be utilized to constantly inform the public of the risk involved in smoking. Smoke cessation clinics should become an integral part of every hospital and health care center. We will show how such smoke cessation programs can lead to highly cost effective results. The third arm of the triad to reduce smoke-related diseases, relates to the low yield cigarette. There is evidence that reduction of the tar yield of a cigarette will, in spite of compensatory smoking habits of such cigarettes, reduce risk of lung and larynx cancer. No such reduction in risk, however, has been shown for cardiovascular disease, indicating different threshold levels for smoke components relating to cancer and cardiovascular diseases. Since tobacco-related diseases continue to make a major impact on the health in both developed and developing nations, increasing attention needs to be given to preventive approaches to tobacco usage.


Age and prior vaccination determine the antibody level in children with primary SARS-CoV-2 Omicron infection.
Protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) reinfection relies on immunity generated after primary infection. However, humoral immunity following primary infection with the Omicron variant is not well understood. We prospectively recruited children <19 years with virologically-confirmed SARS-CoV-2 infection at National Cheng Kung University Hospital from February 2022 to September 2022 during the first wave of Omicron BA.2 outbreak in Taiwan. Serum samples were collected one month after acute infection to measure anti-spike protein receptor binding domain antibody levels and surrogate virus neutralizing antibody (NAb) levels against wild type disease and variants. Of the 164 patients enrolled, most were under 5 years (65.2%) with a diagnosis of upper respiratory tract infection. Children under 6 months with maternal coronavirus disease 2019 (COVID-19) vaccination had higher levels of both anti-SARS-CoV-2 spike antibody (119.0 vs 27.4 U/ml, p < 0.05) and anti-wild type NAb (56.9% vs 27.6% inhibition, p = 0.001) than those without. Children aged 5-12 years with prior vaccination had higher anti-spike antibody, anti-wild type, and anti-Omicron BA.2 NAb levels than those without (all p < 0.05). In previously naïve children without maternal or self-vaccination, those 6 months to 2 years had the highest antibody levels. Multivariable linear regression analysis showed age was the only independent factor associated with antibody level. In our study, children aged 6 months to 2 years have the highest antibody responses to SARS-CoV-2 Omicron variant infection. Age and prior vaccination are the main factors influencing the immunogenicity of SARS-CoV-2 infection.


Effects of a new aminodiphosphonate (aminohydroxybutylidene diphosphonate) in patients with osteolytic lesions from metastases and myelomatosis. Comparison with dichloromethylene diphosphonate.
We have compared in an open trial the clinical and biochemical effects of a new aminodiphosphonate, aminohydroxybutylidene diphosphonate, with those of dichloromethylene diphosphonate, which has been proved effective. The patients presented extensive and symptomatic bone involvement from multiple myeloma, breast cancer, and other metastatic tumors. The treatment consisted of aminohydroxybutylidene diphosphonate, 2.5 mg/d intravenously for five days, or dichloromethylene diphosphonate, 300 mg/d intravenously for seven days, followed by 100 mg/d intramuscularly for ten days. Twelve patients treated with aminohydroxybutylidene diphosphonate and 16 patients treated with dichloromethylene diphosphonate were assessable and were followed up for one to six months. Therapy with aminohydroxybutylidene diphosphonate showed a quicker action in reducing bone pain and reduced significantly more the serum calcium level than did therapy with dichloromethylene diphosphonate. Aminohydroxybutylidene diphosphonate therapy also affected urinary calcium levels and hydroxyproline excretion more markedly than did dichloromethylene diphosphonate, although the differences are not statistically significant. However, the biochemical indexes rebounded more quickly in patients treated with aminohydroxybutylidene diphosphonate, indicating that the loading amount (only 12.5 mg) used in this preliminary study is insufficient to sustain a prolonged effect. The effectiveness and lack of side effects render aminohydroxybutylidene diphosphonate an attractive treatment for malignant bone resorption.


A Novel Regimen for Treating Melanoma: MCL1 Inhibitors and Azacitidine.
Although treatment options for melanoma patients have expanded in recent years with the approval of immunotherapy and targeted therapy, there is still an unmet need for new treatment options for patients that are ineligible for, or resistant to these therapies. BH3 mimetics, drugs that mimic the activity of pro-apoptotic BCL2 family proteins, have recently achieved remarkable success in the clinical setting. The combination of BH3 mimetic ABT-199 (venetoclax) plus azacitidine has shown substantial benefit in treating acute myelogenous leukemia. We evaluated the efficacy of various combinations of BH3 mimetic + azacitidine in fourteen human melanoma cell lines from cutaneous, mucosal, acral and uveal subtypes. Using a combination of cell viability assay, BCL2 family knockdown cell lines, live cell imaging, and sphere formation assay, we found that combining inhibition of MCL1, an anti-apoptotic BCL2 protein, with azacitidine had substantial pro-apoptotic effects in multiple melanoma cell lines. Specifically, this combination reduced cell viability, proliferation, sphere formation, and induced apoptosis. In addition, this combination is highly effective at reducing cell viability in rare mucosal and uveal subtypes. Overall, our data suggest this combination as a promising therapeutic option for some patients with melanoma and should be further explored in clinical trials.


Effect of electrical current on the activities of antimicrobial agents against Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis biofilms.
Bacterial biofilms are resistant to conventional antimicrobial agents. Prior in vitro studies have shown that electrical current (EC) enhances the activities of aminoglycosides, quinolones, and oxytetracycline against Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus epidermidis, Escherichia coli, and Streptococcus gordonii. This phenomenon, known as the bioelectric effect, has been only partially defined. The purpose of this work was to study the in vitro bioelectric effect on the activities of 11 antimicrobial agents representing a variety of different classes against P. aeruginosa, methicillin-resistant Staphylococcus aureus (MRSA), and S. epidermidis. An eight-channel current generator/controller and eight chambers delivering a continuous flow of fresh medium with or without antimicrobial agents and/or EC to biofilm-coated coupons were used. No significant decreases in the numbers of log(10) CFU/cm(2) were seen after exposure to antimicrobial agents alone, with the exception of a 4.57-log-unit reduction for S. epidermidis and trimethoprim-sulfamethoxazole. We detected a statistically significant bioelectric effect when vancomycin plus 2,000 microamperes EC were used against MRSA biofilms (P = 0.04) and when daptomycin and erythromycin were used in combination with 200 or 2,000 microamperes EC against S. epidermidis biofilms (P = 0.02 and 0.0004, respectively). The results of these experiments indicate that the enhancement of the activity of antimicrobial agents against biofilm organisms by EC is not a generalizable phenomenon across microorganisms and antimicrobial agents.


Rosetta predictions in CASP5: successes, failures, and prospects for complete automation.
We describe predictions of the structures of CASP5 targets using Rosetta. The Rosetta fragment insertion protocol was used to generate models for entire target domains without detectable sequence similarity to a protein of known structure and to build long loop insertions (and N-and C-terminal extensions) in cases where a structural template was available. Encouraging results were obtained both for the de novo predictions and for the long loop insertions; we describe here the successes as well as the failures in the context of current efforts to improve the Rosetta method. In particular, de novo predictions failed for large proteins that were incorrectly parsed into domains and for topologically complex (high contact order) proteins with swapping of segments between domains. However, for the remaining targets, at least one of the five submitted models had a long fragment with significant similarity to the native structure. A fully automated version of the CASP5 protocol produced results that were comparable to the human-assisted predictions for most of the targets, suggesting that automated genomic-scale, de novo protein structure prediction may soon be worthwhile. For the three targets where the human-assisted predictions were significantly closer to the native structure, we identify the steps that remain to be automated.


High-yields and extended serum half-life of human interferon alpha2b expressed in tobacco cells as arabinogalactan-protein fusions.
Therapeutic proteins like human interferon alpha2 generally possess short serum half-lives due to their small size, hence rapid renal clearance, and susceptibility to serum proteases. Chemical derivatization, such as addition of polyethylene glycol (PEG) groups overcomes both problems, but at the expense of greatly decreased bioactivity. We describe a new method that yields biologically potent interferon alpha2b (IFNalpha2) in high yields and with increased serum half-life when expressed as arabinogalactan-protein (AGP) chimeras in cultured tobacco cells. Thus IFNalpha2-AGPs targeted for secretion typically gave 350-1400-fold greater secreted yields than the non-glycosylated IFNalpha2 control. The purified AGP domain itself was not immunogenic when injected into mice and only mildly so when injected as a fusion glycoprotein. Importantly, the AGP-IFNalpha2 chimeras showed up to a 13-fold increased in vivo serum half-life while the biological activity remained similar to native IFNalpha2. The use of arabinogalactan glycomodules may provide a general approach to the enhanced production of therapeutic proteins by plants.


Neurotransmitter-Responsive Nanosensors for T2-Weighted Magnetic Resonance Imaging.
Neurotransmitter-sensitive contrast agents for magnetic resonance imaging (MRI) have recently been used for mapping signaling dynamics in live animal brains, but paramagnetic sensors for T1-weighted MRI are usually effective only at micromolar concentrations that themselves perturb neurochemistry. Here we present an alternative molecular architecture for detecting neurotransmitters, using superparamagnetic iron oxide nanoparticles conjugated to tethered neurotransmitter analogs and engineered neurotransmitter binding proteins. Interactions between the nanoparticle conjugates result in clustering that is reversibly disrupted in the presence of neurotransmitter analytes, thus altering T2-weighted MRI signals. We demonstrate this principle using tethered dopamine and serotonin analogs, together with proteins selected for their ability to competitively bind either the analogs or the neurotransmitters themselves. Corresponding sensors for dopamine and serotonin exhibit target-selective relaxivity changes of up to 20%, while also operating below endogenous neurotransmitter concentrations. Semisynthetic magnetic particle sensors thus represent a promising path for minimally perturbative studies of neurochemical analytes.


Prefer what you like? Evaluation and preference of cycling infrastructures in a bicycle simulator.
Previous research on cyclists' route evaluations and preferences already identified influencing factors and relevant evaluation criteria, but studies mostly focused on selected aspects like safety or comfort. This study examined the evaluation of routes more comprehensively considering five evaluation criteria, and further aims to compare the evaluation with the preference of routes. For this, we used the experimental approach of a bicycle simulator. Our participants cycled route segments that varied in certain route characteristics. Each segment was rated in total and on five criteria generated in a previous study, namely Mental Comfort, Interaction, Environment, Ease of Use, and Physical Comfort. At the end, all route segments were ranked according to their quality. Results showed that separated paths were rated the best, while busy footpaths and uphill segments were rated the worst. Interestingly, interacting with pedestrians was described to be more attention-demanding but not as mentally uncomfortable as interacting with motor traffic. The evaluation and preference of routes mostly went hand in hand but differed for the footpath, which was ranked better than it was rated. Results further indicated that gradient has such a strong impact on the physical comfort of a route that it even influenced the overall evaluation. Our findings suggest that the evaluation and preference of routes is influenced by the degree of separation, traffic volume, the type of the road user that the cyclists may need to interact with, but most importantly, it is influenced by the interaction of these three aspects. The five criteria we used can reliably assess the evaluation and preference of routes. They help to differentiate the reason for negative evaluation of routes. This differentiation is crucial to improve cycling routes, as different causes for dissatisfaction among cyclists require different consequences.


Estimating impacts on developing countries of the decrease in U.S. training opportunities for foreign medical graduates.
Between 1973 and 1983, the number of foreign nationals from developing nations who entered the United States for graduate medical education decreased by approximately 90%. Many of those who would have studied in the United States if this decrease had not occurred would have returned home to serve their countries. To estimate the impact of this loss, a survey was conducted in six major cities in Latin America between 1983 and 1989. Selected local medical students interviewed 554 physicians who had returned home after U.S. training and 60 of their classmates who had not trained there. The findings indicate that the returned physicians had given approximately twice as much time to teaching, research, and medical administration as did those who had not left home. The authors maintain that this and related findings show how the curtailment of opportunities for training foreign nationals in the United States is detrimental to both the aspirations of developing nations and the influence of the United States in world affairs.


Differences in infection control and diagnostic measures for multidrug-resistant organisms in the tristate area of France, Germany and Switzerland in 2019 - survey results from the RH(E)IN-CARE network.
Multidrug-resistant organisms (MDROs) are a public health threat. Single-centre interventions, however, are likely to fail in the long term, as patients are commonly transferred between institutions given the economic integration across borders. A transnational approach targeting larger regions is needed to plan overarching sets of interventions. Here, we aim to describe differences in diagnostic and infection prevention and control (IPC) measures in the fight against MDROs. In 2019, we systematically assessed diagnostic algorithms and IPC measures implemented for detection and control of MDROs at three tertiary academic care centres (Freiburg; Strasbourg; Basel). Data were collected using a standardised data collection sheet to be filled in by every centre. Uncertainties were clarified by direct contact via telephone or email with the data supplier. Internal validity was checked by at least two researchers independently filling in the survey. All centres have established a primarily culture-based, rather than a nucleic acid amplification-based approach for detection of MDROs (i.e., vancomycin-resistant Enterococci [VRE], methicillin-resistant Staphylococcus aureus [MRSA], extended-spectrum beta-lactamase-producing Enterobacteriaceae [ESBL], carbapenemase-producing and carbapenem-resistant Gram-negatives [CPGN/CRGN]). IPC measures differed greatly across all centres. High-risk patients are screened for most MDROs on intensive care unit (ICU) admission in all centres; only the French centre is screening all patients admitted to the ICU for VRE, MRSA and ESBL. Patients colonised/infected by MRSA, quinolone-resistant ESBL Klebsiella spp. and CPGN/CRGN are isolated everywhere, whereas patients colonised/infected by VRE and ESBL are usually not isolated in the German centre. In contrast to the French and Swiss centres, the German centre no longer uses isolation measures to control VRE and quinolone-susceptible ESBL. Overall, the French centre is more focused on intercepting MDRO transmission from outside, whereas the German and Swiss centres are more focused on intercepting endemic MDRO transmission. These findings point to important challenges regarding future attempts to standardise IPC measures across borders.   &nbsp.


Markov State Model of Solvent Features Reveals Water Dynamics in Protein-Peptide Binding.
In this work, we investigate the role of solvent in the binding reaction of the p53 transactivation domain (TAD) peptide to its receptor MDM2. Previously, our group generated 831 μs of explicit-solvent aggregate molecular simulation trajectory data for the MDM2-p53 peptide binding reaction using large-scale distributed computing and subsequently built a Markov State Model (MSM) of the binding reaction (Zhou et al. 2017). Here, we perform a tICA analysis and construct an MSM with similar hyperparameters while using only solvent-based structural features. We find a remarkably similar landscape but accelerated implied timescales for the slowest motions. The solvent shells contributing most to the first tICA eigenvector are those centered on Lys24 and Thr18 of the p53 TAD peptide in the range of 3-6 Å. Important solvent shells were visualized to reveal solvation and desolvation transitions along the peptide-protein binding trajectories. Our results provide a solvent-centric view of the hydrophobic effect in action for a realistic peptide-protein binding scenario.


Live face-to-face interaction during fMRI: a new tool for social cognitive neuroscience.
Cooperative social interaction is critical for human social development and learning. Despite the importance of social interaction, previous neuroimaging studies lack two fundamental components of everyday face-to-face interactions: contingent responding and joint attention. In the current studies, functional MRI data were collected while participants interacted with a human experimenter face-to-face via live video feed as they engaged in simple cooperative games. In Experiment 1, participants engaged in a live interaction with the experimenter ("Live") or watched a video of the same interaction ("Recorded"). During the "Live" interaction, as compared to the Recorded conditions, greater activation was seen in brain regions involved in social cognition and reward, including the right temporoparietal junction (rTPJ), anterior cingulate cortex (ACC), right superior temporal sulcus (rSTS), ventral striatum, and amygdala. Experiment 2 isolated joint attention, a critical component of social interaction. Participants either followed the gaze of the live experimenter to a shared target of attention ("Joint Attention") or found the target of attention alone while the experimenter was visible but not sharing attention ("Solo Attention"). The right temporoparietal junction and right posterior STS were differentially recruited during Joint, as compared to Solo, attention. These findings suggest the rpSTS and rTPJ are key regions for both social interaction and joint attention. This method of allowing online, contingent social interactions in the scanner could open up new avenues of research in social cognitive neuroscience, both in typical and atypical populations.


Surveillance of acute rejection in baboon renal transplantation by elevation of interferon-gamma inducible protein-10 and monokine induced by interferon-gamma in urine.
CXCR3 binding chemokines play a key role in recruitment of inflammatory cells into an organ transplant. This study addresses the question of whether urinary excretion of these chemokines correlates with acute rejection in a baboon kidney transplantation model. Seven outbred baboons underwent renal allotransplantation from major histocompatibility complex (MHC)-mismatched donors. The treatment of baboons consisted of anti-CD4 monoclonal antibody (mAb), anti-CD8 mAb, rapamycin, and mycophenolate mofetil (MMF). Urinary levels of interferon-gamma inducible protein-10 (IP-10) and monokine induced by interferon-gamma (Mig) were determined by ELISA. Renal biopsies were examined by immunohistochemical staining for CXCR3 and Mig. Urinary levels of IP-10 and Mig increased significantly in all of the five baboons at the time of acute rejection of renal transplant. The IP-10 and Mig levels did not rise in two nonrejecting baboons. In two baboons, urinary levels of IP-10 and Mig rose before the elevation of the serum creatinine. In renal biopsies, expression of Mig was detected in glomeruli, tubules, and infiltrating cells, and the expression was significantly elevated in biopsies with acute rejection (P<0.01). CXCR3 was constitutively expressed in tubular cells in biopsies derived from both normal grafts and grafts with acute rejection. Whereas the infiltrating cells were increased in the biopsies with acute rejection, the expression of CXCR3 was also significantly higher (P<0.01) in these infiltrating cells compared with those in the normal controls. This study shows an important correlation between urinary excretion of IP-10 and Mig and acute rejection in baboon kidney transplantation.


GST, NAT1, CYP1A1 polymorphisms and risk of esophageal and gastric adenocarcinomas.
Polymorphisms in glutathione-S-transferase (GST), N-acetyltransferase (NAT) 1, and CYP1A1 genes have been suggested as susceptibility factors for esophageal and gastric adenocarcinomas, but have not been consistently linked to elevated risks. In a population-based case-control study, we examined risks in relation to polymorphisms of the following genes: GSTP1; GSTM1; GSTT1; NAT1; and CYP1A1. Histologically confirmed incident cases, ages 30-79, were identified in three US locations. Population controls from the same catchment areas were frequency matched to expected age and sex distributions of esophageal and gastric cardia adenocarcinomas. DNA was extracted from buffy coat for PCR-based assays, with interpretable genotyping results obtained from 209 controls, 67 esophageal adenocarcinomas, 60 gastric cardia adenocarcinomas, and 56 noncardia gastric adenocarcinomas. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated among whites, adjusting for age, sex, and study center. In all histologic subgroups, ORs were somewhat elevated for the GSTP1 Val/Val genotype (versus Ile/Ile), although 95% CIs included 1.00. The respective ORs for esophageal, cardia, and other gastric adenocarcinomas were 1.73 (0.75-4.02), 1.46 (0.57-3.73), and 1.22 (0.48-3.09). No consistent patterns of elevated risk were associated with the null GSTM1 or GSTT1 genotypes, one or two copies of NAT110 or 11 alleles, or CYP1A1 Val/Val or Ile/Val genotypes (versus Ile/Ile). Additional research in larger samples is needed to further assess polymorphisms and their interactions with epidemiologic risk factors, particularly for esophageal adenocarcinoma, which has been increasing markedly in incidence.


Starvation-associated genome restructuring can lead to reproductive isolation in yeast.
Knowledge of the mechanisms that lead to reproductive isolation is essential for understanding population structure and speciation. While several models have been advanced to explain post-mating reproductive isolation, experimental data supporting most are indirect. Laboratory investigations of this phenomenon are typically carried out under benign conditions, which result in low rates of genetic change unlikely to initiate reproductive isolation. Previously, we described an experimental system using the yeast Saccharomyces cerevisiae where starvation served as a proxy to any stress that decreases reproduction and/or survivorship. We showed that novel lineages with restructured genomes quickly emerged in starved populations, and that these survivors were more fit than their ancestors when re-starved. Here we show that certain yeast lineages that survive starvation have become reproductively isolated from their ancestor. We further demonstrate that reproductive isolation arises from genomic rearrangements, whose frequency in starving yeast is several orders of magnitude greater than an unstarved control. By contrast, the frequency of point mutations is less than 2-fold greater. In a particular case, we observe that a starved lineage becomes reproductively isolated as a direct result of the stress-related accumulation of a single chromosome. We recapitulate this result by demonstrating that introducing an extra copy of one or several chromosomes into naïve, i.e. unstarved, yeast significantly diminishes their fertility. This type of reproductive barrier, whether arising spontaneously or via genetic manipulation, can be removed by making a lineage euploid for the altered chromosomes. Our model provides direct genetic evidence that reproductive isolation can arise frequently in stressed populations via genome restructuring without the precondition of geographic isolation.


Antineoplastic activity of zoledronic acid and denosumab.
Cancer patients suffer from cancer-induced bone pain, hypercalcemia, and reduced quality of life caused by pathological fractures. Many of these complications related to cancer can be treated, or at least controlled, using new anticancer agents. Recently, two agents used initially to treat osteoporosis demonstrated direct and indirect anticancer activity. In this review, we summarize current knowledge about direct and indirect anticancer activity of zoledronic acid (a third-generation bisphosphonate), and denosumab antibody against RANKL. Zoledronic acid influences the proliferation and viability of tumor cells in vitro, and effectively reduces tumor burden, tumor-induced pain, and tumor growth in vivo. Denosumab is a fully human monoclonal antibody preventing the binding of RANKL to its receptor on osteoclasts' membrane, and through this mechanism inhibits the resorption of the bone. Furthermore, this agent demonstrates direct anticancer activity through the RANKL signaling pathway. Because of these features both drugs may gain broader application for the treatment of cancer patients. However, further pre-clinical and clinical evaluation is needed for both agents to fully assess the antineoplastic mechanisms of activity of both agents.


Comparison of conformal radiation therapy techniques within the dynamic radiotherapy project 'Dynarad'.
The objective of the dynamic radiotherapy project 'Dynarad' within the European Community has been to compare and grade treatment techniques that are currently applied or being developed at the participating institutions. Cervical cancer was selected as the tumour site on the grounds that the involved organs at risk, mainly the rectum and the bladder, are very close to the tumour and partly located inside the internal target volume. In this work, a solid phantom simulating the pelvic anatomy was used by institutions in Belgium, France, Greece, Holland, Italy, Sweden and the United Kingdom. The results were evaluated using both biological and physical criteria. The main purpose of this parallel evaluation is to test the value of biological and physical evaluations in comparing treatment techniques. It is demonstrated that the biological objective functions allow a much higher conformality and a more clinically relevant scoring of the outcome. Often external beam treatment techniques have to be combined with intracavitary therapy to give clinically acceptable results. However, recent developments can reduce or even eliminate this need by delivering more conformal dose distributions using intensity modulated external dose delivery. In these cases the reliability of the patient set-up procedure becomes critical for the effectiveness of the treatment.


The development of activatable lytic peptides for targeting triple negative breast cancer.
Cytolytic peptides are an emerging class of promising cancer therapeutics shown to overcome drug resistance. They eliminate cancer cells via disruption of the phospholipid bilayer of cell membranes, a mechanism that differentiates it from traditional treatments. However, applications of lytic peptides via systematic administration are hampered by nonspecific toxicity. Here, we describe activatable, masked lytic peptides that are conjugated with anionic peptides via a cleavable linker sensitive to matrix metalloproteinases (Ac-w-βA-e8-XPLG*LAG-klUklUkklUklUk-NH2; lower case letters in the sequences represent D-amino-acids, U=Aib, α-aminoisobutyric acid, *cleavage site). The peptides were activated upon being introduced into the triple negative breast cancer cell line MDA-MB-231, which overexpresses secreted matrix metalloproteinases, to selectively cleave the peptide linker. Our results indicate that the activatable design could be applied to improve the targeting ability of lytic peptides.


Fc(epsilon)RI as a paradigm for a lipid raft-dependent receptor in hematopoietic cells.
Lipid domains or rafts are currently embraced by immunologists as critical participants in receptor-mediated signaling events occurring at the plasma membrane. This view of membrane heterogeneity and its functional importance is supported by many years of different experimental approaches. We can now refine our investigations, moving beyond the simple models to ask more detailed questions about structural properties and mechanistic interactions. As highlighted for the IgE receptor (Fc(epsilon)RI), new information about initial engagement with src family kinases, cytoskeletal regulation, and coupling with downstream signaling is beginning to emerge.


Prevalence of Pulmonary Embolism in Patients Presenting to the Emergency Department for Syncope.
The Research to Practice column is intended to improve the research critique skills of the advanced practice registered nurse and the emergency nurse (RN) and to assist with the translation of research into practice. For each column, a topic and a research study are selected. The research article is then reviewed and critiqued, and the findings are discussed in relation to a patient scenario. In this column, we examine the findings of P. from their article, titled "Prevalence of Pulmonary Embolism Among Patients Hospitalized for Syncope."


Novel Missense LCAT Gene Mutation Associated with an Atypical Phenotype of Familial LCAT Deficiency in Two Portuguese Brothers.
Familial lecithin-cholesterol acyltransferase deficiency (FLD) is a rare recessive disorder of cholesterol metabolism, caused by loss-of-function mutations in the human LCAT gene, leading to alterations in the lipid/lipoprotein profile, with extremely low HDL levels.The classical FLD phenotype is characterized by diffuse corneal opacification, haemolytic anaemia and proteinuric chronic kidney disease (CKD); an incomplete form, only affecting the corneas, has been reported in a few families worldwide.We describe an intermediate phenotype of LCAT deficiency, with CKD preceding the development of corneal clouding, in two Portuguese brothers apparently homozygous for a novel missense LCAT gene mutation. The atypical phenotype, the diagnosis of membranous nephropathy in the proband's native kidney biopsy, the late-onset and delayed recognition of the corneal opacification, the co-segregation with Gilbert syndrome and the late recurrence of the primary disease in kidney allograft all contributed to obscure the diagnosis of an LCAT deficiency syndrome for many years.A major teaching point is that on standard light microscopy examination the kidney biopsies of patients with LCAT deficiency with residual enzyme activity may not show significant vacuolization and may be misdiagnosed as membranous nephropathy. The cases of these two patients also illustrate the importance of performing detailed physical examination in young adults presenting with proteinuric CKD, as the most important clue to the diagnosis of FLD is in the eyes.


Phase I study of bryostatin 1 in patients with relapsed non-Hodgkin's lymphoma and chronic lymphocytic leukemia.
To define, in a phase I study in relapsed non-Hodgkin's lymphoma (NHL) and chronic lymphocytic leukemia (CLL), the maximum-tolerated dose (MTD), major toxicities, and possible antitumor activity of bryostatin 1, a macrocyclic lactone. Bryostatin 1 was delivered by 72-hour continuous infusion every 2 weeks to patients with relapsed NHL or CLL, at doses that ranged from 12 microg/m2 to 180 microg/m2 per course. Correlative investigations included evaluations of total protein kinase C (PKC) in peripheral blood and lymphoid differentiation in patient tumor tissue. Twenty-nine patients were treated, including three patients with CLL and 26 with NHL. Generalized myalgia was the dose-limiting toxicity (DLT) and occurred in two of three patients treated with bryostatin 1 at 180 microg/m2 per course. Myalgias were dose-related and cumulative, and often started in the thighs and calves, improved with activity, were somewhat responsive to analgesics, and often took weeks to resolve once taken off study. Six patients were treated at the MTD of 120 microg/m2 per course. Myalgia, headache, and fatigue were common. Hematologic toxicity was uncommon. Total cumulative doses of bryostatin 1 up to 1,134 microg/m2 have been administered without untoward toxicity. Eleven patients achieved stable disease for 2 to 19 months. An in vitro assay for total PKC evaluation in patient peripheral-blood samples demonstrated activation within the first 2 hours with subsequent downregulation by 24 hours, which was maintained throughout the duration of the 72-hour infusion. This phase I study defined the MTD and recommended phase II dose of bryostatin 1, when administered over 72 hours every 2 weeks, to be 120 microg/m2 (40 microg/m2/d for 3 days). Generalized myalgia was the DLT. Future studies will define the precise activity of bryostatin 1 in subsets of patients with lymphoproliferative malignancies and its efficacy in combination with other agents.


Attenuation characteristics of the fundamental modes that propagate in buried iron water pipes.
The attenuation of the fundamental non-torsional modes that propagate down buried iron water pipes has been studied. The mode shapes, mode attenuation due to leakage into the surrounding medium and the scattering of the modes as they interact with pipe joints and fittings have been investigated. In the low frequency region the mode predicted to dominate over significant propagation distances approximates a plane wave in the water within the pipe. The established acoustic technique used to locate leaks in buried iron water pipes assumes that leak noise propagates as a single non-dispersive mode at a velocity related to the low frequency asymptote of this water borne mode. Experiments have been conducted on buried water mains at test sites in the UK to verify the attenuation and velocity dispersion predictions.


Restless legs syndrome and multiple sclerosis: a Brazilian multicenter study and meta-analysis of the literature.
The increased prevalence of restless legs syndrome (RLS) in multiple sclerosis (MS) has recently been the subject of a few papers. The present study investigated the prevalence of RLS symptoms in MS patients and in controls in four Brazilian cities. Additionally, a systematic review and meta-analysis of the literature was carried out on the subject of RLS-MS. MS patients and controls were investigated regarding the presence of the four typical symptoms of RLS. A questionnaire assessing RLS severity was also used for patients and controls presenting the four RLS symptoms criteria. The systematic review and meta-analysis on the subject were carried out according to the strict international criteria. In the present report, the RLS-MS association was confirmed as being more than fortuitous in Brazilian MS patients, in a multicenter case-control study. MS patients also presented RLS symptoms of greater severity than did the control population. A systematic review and meta-analysis of the literature showed that MS patients had a fourfold higher chance of presenting RLS than did the controls. Although underlying mechanisms to explain the association RLS-MS are still a matter of discussion, there is a clear association of these two neurological conditions.


Association between Hepatitis B Surface Antigen Levels and the Risk of Hepatocellular Carcinoma in Patients with Chronic Hepatitis B Infection: Systematic Review and Meta-Analysis.
Background: The role of hepatitis B surface antigen (HBsAg) levels in predicting the risk of developing hepatocellular carcinoma (HCC) has remained unclear. The aim of this study was to obtain the most up-to-date estimated measure of the association between HBsAg levels and the development of HCC in patients. Methods: We performed a systematic review by searching for relevant studies on PubMed, Scopus, ProQuest and the Cochrane Central Register of Controlled Trials from January 2002 to November 2017. We presented the effects of HBsAg levels at each cut-off value as the odds ratios (ORs) at 95% confidence interval (CI). We also investigated HCC and its potential risk factors including HBeAg, and HBV DNA. We registered our protocol with the International Prospective Register of Systematic Reviews (PROSPERO) with the registration number CRD42018081138. Results: We selected 10 studies representing 12 541 cases. At the 100 IU/ml cut-off, the OR for HCC at the high HBsAg level versus the low level was 4.99 (95% CI, 3.01–8.29) with high inconsistency (I2=79%). At the 1,000 IU/ml threshold, the pooled OR for HCC at the high HBsAg versus the low level was 2.46 (95% CI, 2.15–2.83) with low variance. We also found correlations between the risk of HCC and male gender (OR=2.12), hepatitis B e-antigen positivity (OR=2.99), or hepatitis B (HBV) viral load ≥ 2,000 IU/ml (OR=4.37). Conclusion: Our study revealed that HBsAg levels ≥ 100 IU/ml, and notably >1,000 IU/ ml, are associated with an increased risk of HCC development.


Climate adaptation in the public health sector in Africa: Evidence from United Nations Framework Convention on Climate Change National Communications.
Climate change has potential to affect human health in various ways. Extreme temperatures and cold both result in deaths, while the changing habitats favouring the breeding of vectors could result in the spread of diseases such as malaria, cholera and typhus. This article reviews climate change adaptation measures in the African public health sector. The evidence is drawn from National Communications of 21 countries as submitted to the United Nations Framework Convention on Climate Change (UNFCCC). This article combines the literature review and grounded theory approaches with data obtained from the UNFCCC National Communications. Among key adaptation measures emerging from the work are weather-based forecasting and early warning systems, public education and awareness, putting in place appropriate policies, surveillance, research and monitoring as well as improving public health infrastructure and technology. The study recommends that African nations should commit to address health impacts of climate change through the implementation of appropriate adaptation measures.