Atypical postinfectious glomerulonephritis is associated with abnormalities in the alternative pathway of complement.
Postinfectious glomerulonephritis is a common disorder that develops following an infection. In the majority of cases, there is complete recovery of renal function within a few days to weeks following resolution of the infection. In a small percentage of patients, however, the glomerulonephritis takes longer to resolve, resulting in persistent hematuria and proteinuria, or even progression to end-stage kidney disease. In some cases of persistent hematuria and proteinuria, kidney biopsies show findings of a postinfectious glomerulonephritis even in the absence of any evidence of a preceding infection. The cause of such 'atypical' postinfectious glomerulonephritis, with or without evidence of preceding infection, is unknown. Here we show that most patients diagnosed with this 'atypical' postinfectious glomerulonephritis have an underlying defect in the regulation of the alternative pathway of complement. These defects include mutations in complement-regulating proteins and antibodies to the C3 convertase known as C3 nephritic factors. As a result, the activated alternative pathway is not brought under control even after resolution of the infection. Hence, the sequela is continual glomerular deposition of complement factors with resultant inflammation and development of an 'atypical' postinfectious glomerulonephritis.


Gradual rebalancing of Medicaid long-term services and supports saves money and serves more people, statistical model shows.
States are shifting Medicaid spending on long-term services and supports from institutional to home and community-based services, a process known as rebalancing. Using fifteen years of state expenditure data, a statistical model was developed to assess the effect of rebalancing on overall spending for long-term services and supports. The model indicates that spending is affected by the way rebalancing is implemented: Gradual rebalancing, by roughly two percentage points annually, can reduce spending by about 15 percent over ten years. More rapid rebalancing can save money, break even, or increase spending, depending on the pace and program specifics. Cuts to home and community-based services that hinder rebalancing are likely to increase, not decrease, overall spending on long-term services and supports as people who were receiving these services shift into nursing homes. Because many states continue to experience budget crises, policy makers must think carefully before altering spending patterns for long-term services and supports and adopt strategies that particular states have used to successfully reduce overall spending, such as gradually shifting expenditures toward home and community-based waiver programs.


Chronic hypoxia induces proliferation of cultured mesangial cells: role of calcium and protein kinase C.
The effect of hypoxia on the proliferation of cultured rat mesangial cells was examined. To evaluate the underlying signaling mechanisms, the roles of intracellular calcium ([Ca2+]i) and protein kinase C (PKC) were determined. Quiescent cultures were exposed to hypoxia (3% O2) or normoxia (18% O2), and [3H]thymidine incorporation, cell number, [Ca2+]i, and PKC were assessed. Mesangial cells exposed to 28 h of hypoxia exhibited a significant increase in [3H]thymidine incorporation followed by a significant increase in cell number at 72 h in comparison with respective normoxic controls. Hypoxia induced a biphasic activation of PKC, reflected by translocation of the enzyme activity from cytosol to membrane at 1 h, a return to baseline at 4 and 8 h, with subsequent reactivation from 16 to 48 h. In addition, hypoxia-induced proliferation was prevented by a PKC inhibitor 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7). Cells exposed to hypoxia produced progressive increases in resting [Ca2+]i from 15 to 60 min which remain sustained up to 24 h of examination. Verapamil significantly prevented the hypoxia-induced proliferation, and both verapamil treatment and incubations in a calcium-free medium for 1 h blocked the hypoxia-induced stimulation of [Ca2+]i as well as PKC. These results provide the first in vitro evidence that chronic hypoxia induces proliferation of cultured glomerular mesangial cells, which is mediated by the stimulation of [Ca2+]i and the subsequent activation of PKC.


Inhibition of neointimal formation by trans-resveratrol: role of phosphatidyl inositol 3-kinase-dependent Nrf2 activation in heme oxygenase-1 induction.
Neointima, defined as abnormal growth of the intimal layer of blood vessels, is believed to be a critical event in the development of vascular occlusive disease. Although resveratrol's inhibitory effects on proliferation and migration of vascular smooth muscle cells has been reported, its activity on neointimal formation is still unclear. Oral administration of trans-resveratrol significantly suppressed intimal hyperplasia in a wire-injured femoral artery mouse model. In cultured vascular smooth muscle cells, trans-resveratrol inhibited platelet-derived growth factor-stimulated DNA synthesis and cell proliferation with down-regulation of cyclin D and pRB. Moreover, platelet-derived growth factor-induced production of reactive oxygen species was inhibited by trans-resveratrol and the compound induced heme oxygenase-1 (HO-1). The anti-proliferative activity of trans-resveratrol was reversed by an HO-1 inhibitor, ZnPPIX. Subcellular fractionation and reporter gene analyses revealed that trans-resveratrol increased the level of nuclear Nrf2 and antioxidant response element reporter activity, and that these were essential for the induction of HO-1. Trans-resveratrol also enhanced the activities of phosphatidyl inositol 3-kinase and extracellular signal regulated kinase, and phosphatidyl inositol 3-kinase was required for Nrf2/antioxidant response element-dependent HO-1 induction. These data have significant implications for the elucidation of the pharmacological mechanism by which trans-resveratrol prevents vascular occlusive diseases.


Muscle pressor reflex: potential role of vanilloid type 1 receptor and acid-sensing ion channel.
Reflex cardiovascular responses to muscle contraction are mediated by mechanical and metabolic stimulation of thin muscle afferent fibers. Metabolic stimulants and receptors involved in responses are uncertain. Capsaicin depolarizes thin sensory afferent nerves that have vanilloid type 1 receptors (VR1). Among potential endogenous ligands of thin fibers, H+ has been suggested as a metabolite mediating the reflex muscle response as well as a potential stimulant of VR1. It has also been suggested that acid-sensing ion channels (ASIC) mediate H+, evoking afferent nerve excitation. We have examined the roles of VR1 and ASIC in mediating cardiovascular reflex responses to acid stimulation of muscle afferents in a rat model. In anesthetized rats, injections of capsaicin into the arterial blood supply of triceps surae muscles evoked a biphasic response (n = 6). An initial fall in mean arterial pressure (from baseline of 95.8 +/- 9.5 to 70.4 +/- 4.5 mmHg, P < 0.05 vs. baseline) was followed by an increase (to 131.6 +/- 11.3 mmHg, P < 0.05 vs. baseline). Anandamide (an endogenous substance that activates VR1) induced the same change in blood pressure as did capsaicin. The pressor (but not depressor) component of the response was blocked by capsazepine (a VR1 antagonist) and section of afferent nerves. In decerebrate rats (n = 8), H+ evoked a pressor response that was not blocked by capsazepine but was attenuated by amiloride (an ASIC blocker). In rats (n = 12) pretreated with resiniferatoxin to destroy muscle afferents containing VR1, capsaicin and H+ responses were blunted. We conclude that H+ stimulates ASIC, evoking the reflex response, and that ASIC are likely to be frequently found on afferents containing VR1. The data also suggest that VR1 and ASIC may play a role in processing of muscle afferent signals, evoking the muscle pressor reflex.


Age and growth of the midwater crocodile shark Pseudocarcharias kamoharai.
Age and growth were analysed on the basis of 372 vertebrae from specimens of the crocodile shark Pseudocarcharias kamoharai (66·0-122·0 cm, total length, LT ) collected in the south-western Atlantic Ocean. Centrum edge analysis suggested the first four months of the year as the period of band completion, leading to acceptance of free-living bands as annual. A pre-birth ring is formed in embryos >31·0 cm LT , whereas the birthmark is deposited in newborns of 46·1 ± 1·1 cm LT (mean ± s.d.). Growth was parameterized using seven models and the AIC was used for selection of the most plausible model. The von Bertalanffy growth model received the greatest support from the data, providing the following growth parameters for combined sexes: L∞ = 129·2 cm, k = 0·137 year(-1) and t0 = -3·9 years. No significant differences were detected in growth by sex, but significant differences in age composition by sex were found in the sample. Overall age ranged from 2 to >13 years in females and from 1 to 8 years in males. Males achieved first maturity at 3·1 years and females at 5·1 years, with pregnancy appearing at 4·8 years. Similar to other lamnoid species, P. kamoharai is relatively fast growing.


Current densities in a 2 mm resolution anatomically realistic model of the body induced by low frequency electric fields.
Current density distributions in a fine resolution (2 mm) anatomically realistic voxel model of the human body have been calculated for uniform, low frequency vertically aligned electric fields for a body grounded and isolated from 50 Hz to 10 MHz. The voxel phantom NORMAN is used which has a height of 1.76 m and a mass of 73 kg. There are 8.3 million voxels in the body differentiated into 37 tissue types. Both finite-difference potential and time-domain methods were used. Results are presented for the current density averaged over 1 cm2 in muscle, heart, brain and retina. Electric field values required to reach the NRPB and ICNIRP basic restrictions on current density are derived and compared with the external field guidelines from these standards.


Amplified spontaneous emission and optical gain measurements from pyrromethene 567--doped polymer waveguides and quasi-waveguides.
Amplified spontaneous emission from planar waveguides and quasi-waveguides based on Pyrromethene 567-doped poly(methyl methacrylate) thin films deposited onto quartz and glass substrates is investigated. Films with different thickness were prepared and pumped optically at 532 nm with pulses of up to 8 MW/cm(2). Pump thresholds for the onset of ASE emission, optical gains and losses were assessed. Net gain coefficients were estimated by fitting the data provided by variable stripe length measurements with a theoretical expression which takes into account saturation. In this way, net gain coefficients of up to 56 +/- 9 cm(-1) at a pump intensity of 5.3 MW/cm(2) for quasi-waveguides and up to 20.6 +/- 2.7 cm(-1) at a pump intensity of 3.4 MW/cm2 for waveguides, were obtained. Loss coefficients in the waveguides were estimated to be 3.8 +/- 0.4 cm-1 and 6.1 +/- 1.3 cm(-1) for 15 microm and 5 microm thick films, respectively. The results obtained seem to indicate a stronger self-mode-restriction capability in the quasiwaveguides than in conventional total internal-reflection waveguides.


Environmental lead exposure increases micronuclei in children.
The objective of this pilot study was to investigate the contribution of environmental exposures to lead in the development of cytogenetic damage detected as the frequency of micronuclei (MN) in children. The other aim was to apply the MN assay in combination with fluorescence in situ hybridization (FISH) using a pan-centromeric chromosome probe to elucidate the formation mechanism of induced MN. The examined population was composed of 9-year-old children (n = 92), living in the region where non-ferrous ores are extracted and processed. The non-exposed group consisted of 49 children of the same age from an unexposed recreational area. Exposure to lead was assessed by determination of lead concentrations in blood (PbB) by atomic absorption spectrophotometry, whereas the level of selenium (Se) in serum was detected by using graphite furnace atomic-absorption spectrometry. The frequency of MN was determined by the cytokinesis-block MN assay and fluorescence in situ hybridization performed using a specific pan-centromeric probe. Environmental exposure to lead resulted in significantly increased levels of PbB (5.29 +/- 2.09 versus 3.45 +/- 1.20 microg/dl in controls), although the average level was much below the value of the biological exposure limit = 10 microg/dl. A negative correlation between lead in blood and Se in serum concentrations (P = 0.006) was found for the pooled study population. The results showed a significant difference (P < 0.0001) in the level of MN between the exposed and control group (standard MN test: 2.96 +/- 2.36 versus 1.16 +/- 1.28; FISH technique: 3.57 +/- 3.02 versus 1.43 +/- 1.69, respectively). The frequencies of both centromere-positive (C+MN) and centromere-negative (C-MN) micronuclei were significantly increased in exposed children; however, the contribution of C+MN in the total number of MN in peripheral blood lymphocytes of exposed children was significantly higher than in the controls what may suggest a pro-aneugenic effect of the exposure to lead. The results of multiple regression analysis indicated that the exposure to lead was an important factor affecting the increase in MN frequency what was confirmed by significant correlation between the PbB and MN levels. In conclusion, our results suggest that the exposure to lead may be associated with an increased frequency of MN, especially of C+MN; however, the influence of other factors (e.g. vitamins and minerals in the diet) cannot be excluded.


Talonavicular joint fixation using augmenting naviculocalcaneal screw in modified double hindfoot arthrodesis.
Triple arthrodesis and combined talonavicular/subtalar arthrodesis are procedures for hindfoot disorders. Achieving talonavicular joint fusion can be problematic. We hypothesized that talonavicular joint fixation bending stiffness could be increased by adding a fully-threaded screw inserted through the lateral navicular into the calcaneus. We used ten fresh-frozen cadaver hindfeet. The calcaneus and talus were immobilized, and talonavicular joint stripped of cartilage. Two partially-threaded 4.5-mm cannulated screws were placed across the talonavicular joint, one traversing the central third of the navicular and a second through the medial third of the navicular. We quantified talonavicular fixation bending stiffness and angulation in response to 4N-m bending moments applied to the navicular in four directions: plantar-to-dorsal (P-D), medial-to-lateral (M-L), dorsal-to-plantar (D-P), and lateral-to-medial (L-M). An augmenting fully-threaded 4.5 mm cannulated screw was then placed through the lateral third of the navicular into the calcaneus and the specimens retested. The mean bending stiffness in each direction increased significantly after addition of the third screw. The stiffness increase in each direction was: P-D 92%, D-P 39%, L-M 108%, and M-L 56%. Correspondingly, mean talonavicular angulation significantly decreased in each direction: P-D 43%, D-P 28%, L-M 47%, and M-L 34%. A third, fully-threaded screw from the lateral navicular into the calcaneus significantly increases talonavicular bending stiffness and decreases angulation. The increased resistance to arthrodesis site motion afforded by the addition of a naviculocalcaneal screw may improve the likelihood of achieving successful talonavicular fusion in combined talonavicular/subtalar arthrodesis.


Flemish and Dutch mutations in amyloid beta precursor protein have different effects on amyloid beta secretion.
Mutations in the amyloid beta precursor protein (APP) gene cosegregate with autosomal dominant Alzheimer disease (AD). Brain pathology of AD is characterized by amyloid deposition in senile plaques and by neurofibrillary tangles. Amyloid deposits in AD brains consist of amyloid beta (A beta), a 4-kDa proteolytic product of APP. In contrast, two other mutations in APP, the Flemish APP692 and Dutch APP693 mutations, are associated with autosomal dominant cerebral hemorrhages due to congophilic amyloid angiopathy (CAA) in the presence or absence of AD pathology, respectively. Both mutations are located within A beta near the constitutive cleavage site. While a common effect of AD-linked mutations is to elevate A beta 42 extracellular concentrations, not much is known about the effect of APP692 and APP693. Here we provide evidence that APP692 and APP693 have a different effect on A beta secretion as determined by cDNA transfection experiments. While APP692 upregulates both A beta 40 and A beta 42 secretion, APP693 does not. These data corroborate with previous findings that increased A beta secretion and particularly of A beta 42, is specific for AD pathology.


Nuclear medicine in the acute clinical setting: indications, imaging findings, and potential pitfalls.
Nuclear medicine imaging provides valuable functional information that complements information obtained with anatomic imaging techniques in the evaluation of patients with specific acute clinical manifestations. Nuclear medicine studies are most often used in conjunction with other imaging modalities and as a problem-solving tool. Under certain circumstances a nuclear medicine study may be indicated as the first-line imaging modality, as in the case of renal scintigraphy for transplant dysfunction in the early postoperative period. Nuclear imaging may be preferred when a conventional first-line study is contraindicated or when it is important to minimize radiation exposure. The portability of nuclear imaging offers particular advantages for the evaluation of critically ill patients whose clinical condition is unstable and who cannot be safely transported out of the intensive care unit. The ability to visualize physiologic and pathophysiologic processes over relatively long time periods without adding to the patient's radiation exposure contributes to the high diagnostic sensitivity of several types of nuclear medicine studies. Viewing the acquired images in the cine mode adds to the value of these studies for diagnosing and characterizing dynamic abnormalities such as intermittent internal bleeding and bile or urine leakage. In this pictorial review, the spectrum of nuclear medicine studies commonly performed in the acute care setting is reviewed according to body systems and organs, with detailed descriptions of the indications, technical considerations, findings, and potential pitfalls of each type of study. Supplemental material available at http://radiographics.rsna.org/lookup/suppl/doi:10.1148/rg.332125098/-/DC1.


aKmerBroom: Ancient oral DNA decontamination using Bloom filters on k-mer sets.
Dental calculus samples are modeled as a mixture of DNA coming from dental plaque and contaminants. Current computational decontamination methods such as Recentrifuge and DeconSeq require either a reference database or sequenced negative controls, and therefore have limited use cases. We present a reference-free decontamination tool tailored for the removal of contaminant DNA of ancient oral sample called aKmerBroom. Our tool builds a Bloom filter of known ancient and modern oral k-mers, then scans an input set of ancient metagenomic reads using multiple passes to iteratively retain reads likely to be of oral origin. On synthetic data, aKmerBroom achieves over 89.53% sensitivity and 94.00% specificity. On real datasets, aKmerBroom shows higher read retainment (+60% on average) than other methods. We anticipate aKmerBroom will be a valuable tool for the processing of ancient oral samples as it will prevent contaminated datasets from being completely discarded in downstream analyses.


Molecular epitopes of the ankyrin-spectrin interaction.
Isoforms of ankyrin and its binding partner spectrin are responsible for a number of interactions in a variety of human cells. Conflicting evidence, however, had identified two different, non-overlapping human erythroid ankyrin subdomains, Zu5 and 272, as the minimum binding region for beta-spectrin. Complementary studies on the ankyrin-binding domain of spectrin have been somewhat more conclusive yet have not presented binding in terms of well-phased, integral numbers of spectrin repeats. Thus, the objective of this study was to clearly define and characterize the minimal ankyrin-spectrin binding epitopes. Circular dichroism (CD) wavelength spectra of the aforementioned ankyrin subdomains show that these fragments are 30-60% unstructured. In contrast, human erythroid beta-spectrin repeats 13, 14, 15, and 16 (prepared in all combinations of two adjacent repeats) demonstrated proper folding and stability as determined by CD and tryptophan wavelength and heat denaturation scans. Native polyacrylamide gel electrophoresis (PAGE) gel shifts as well as affinity pull-down assays implicated Zu5 and beta-spectrin repeats 14-15 as the minimum binding epitopes. These results were confirmed by analytical ultracentrifugation to sedimentation equilibrium by which a 1:1 complex was obtained if and only if Zu5 was mixed with beta-spectrin constructs containing repeats 14 and 15 in tandem. Surface plasmon resonance yielded a K D of 15.2 nM for binding of beta-spectrin fragments to the ankyrin subdomain Zu5, accounting for all of the binding observed between the intact molecules. Collectively, these results show the 14th and 15th beta-spectrin repeats comprise the minimal, phased region of beta-spectrin, which binds ankyrin at the Zu5 subdomain with high affinity.


Incidence of cleft lip and palate and risks of additional malformations.
Children with cleft lip and/or palate (n = 251) born between 1991 and 1995 in the county of Stockholm, Sweden, were studied with reference to incidence and rate ratios (RRs) of different types of clefts, gender, birth weight, mother's age, and length of pregnancy. Children who had clefts and additional malformations were compared with children who had clefts but no additional malformations. The incidence of clefts was 2.0/1000 live births, and it was higher among males than among females. The RR, an index of relative risk, was 1.58. The main groups, children with isolated cleft lip, children with cleft lip and palate, and children with isolated cleft palate, showed similar incidence values (0.6-0.7/1000 live births). Children with bilateral clefts had an incidence of 0.3/1000 live births. Additional malformations were found in approximately every sixth newborn with a cleft when children with Robin sequence were excluded. There was a tendency for newborns with bilateral clefts to have additional malformations (RR = 1.36; confidence interval = 0.74-2.49). Children with clefts and additional malformations had lower birth weight and were born earlier than children with clefts only. Preterm cleft children with low birth weight should be screened for the presence of other birth defects.


Angiographic evaluation of renal allografts.
In a material of 128 kidneys transplanted to 112 patients 361 angiographies have been carried out in early, as well as in late course. The method is a valuable tool in the diagnosis of rejection, particularly in the early posttransplantation period when the kidney does not function due to tubular necrosis. Angiography also has prognostic significance, and can therefore be of value when deciding if a graft should be removed or if continuous efforts should be made to save it.


Electrophysiological effects of nitric oxide on spontaneous activity of rabbit atrioventricular node cells.
The electrophysiological effects of nitric oxide (NO) on spontaneous activity of rabbit atrioventricular (AV) node cells were examined using intracellular microelectrode technique. The results obtained are as follows. (1) NO donors sodium nitroprusside (SNP, 1~1000 micromol/L) and 3-morpholinosydnonimine (SIN-1, 100, 1000 micromol/L) decreased the amplitude of action potential (APA), rate of spontaneous firing (RSF), velocity of diastolic (phase 4) depolarization (VDD), and maximal rate of depolarization (V(max)) in a concentration-dependent manner. (2) Pretreatment with L-type calcium channel agonist Bay K8644 (0.25 micromol/L) completely reversed the effects of SNP (100 micromol/L) on AV node cells. (3) Elevation of Ca(2+) concentration (5 mmol/L) in superfusate antagonized the effects of SNP on AV node cells. (4) Perfusion with Ca(2+)-free K-H solution, completely abolished the effects of SNP on AV node cells. (5) Application of methylene blue (50 micromol/L), a guanylyl cyclase inhibitor, failed to abolish the inhibitory effects of SNP (100 micromol/L). All these results suggest that NO exerts a negative effect on spontaneous activity of AV node cells in rabbits. These effects are likely due to reduction in calcium influx via a cGMP-independent mechanism.


Decontamination of human xenotransplantable tumor with mouse hepatitis virus by implantation in nude rat: a case report.
A human tumor xenograft contaminated with mouse hepatitis virus (MHV) was implanted in a nude rat in order to decontaminate the tumor line. The decontamination failed in the first trial, but succeeded in the second trial. The difference between the two trials was the duration of implantation of the tumor in the nude rat, i.e., 12 days in the first and 24 days in the second trial. Duration of implantation might be a factor in the decontamination of transplantable tumors infected with MHV by passaging in the nude rat.


The presence of plasma proteins facilitates the uptake of 125I-thrombin by the rabbit thoracic aorta endothelium in vitro.
Various purified proteins, protein derivatives and two polysaccharides were added individually to a physiological medium in order to effect uptake of 125I-thrombin by the rabbit aorta endothelium. Over a wide range of concentration (0.004-40 mg/ml), the presence of either purified rabbit or bovine albumin during thrombin uptake encouraged an increase (70-110%) in 125I-thrombin binding by the endothelium and subendothelium compared to uptake by aorta segments in the absence of added protein. Pretreatment of aorta segments with albumin before incubation with 125I-thrombin in the absence of albumin did not encourage thrombin uptake to the same extent as having 125I-thrombin and albumin together. Purified human transferrin, rabbit IgG, chicken ovalbumin or denatured bovine casein could replace albumin to produce a similar enhancement of thrombin uptake. Replacing active concentrations of albumin by either reduced-carboxymethylated albumin, defatted albumin, plasmin-treated or thermolysin-treated albumin also caused an increase (50-130%) in thrombin binding, whereas replacement by acid-hydrolysed albumin or with polyglutamic acid was either ineffective or even inhibitory. Lysine-modified or arginine-modified albumins caused a small enhancement (14-32%) and no enhancement of thrombin uptake, respectively. Dextran, at low concentration (0.04-0.4 mg/ml) did not influence thrombin uptake, and at higher concentration (4-40 mg/ml) caused a decrease in uptake by both the endothelium and subendothelial layers. Low concentration of dextran sulphate inhibited thrombin uptake to 20-30% of control values. These data express the importance of accompanying protein in the response of the vascular endothelium during binding of thrombin. The possibility that other protein-cell interactions may be similarly influenced by macromolecular solutes is also discussed.


Evaluation of an automated erythrocyte sedimentation rate analyzer as compared to the Westergren manual method in measurement of erythrocyte sedimentation rate.
Monitor 100® (Electa Lab, Italy) is a newly developed automated method for measurement of erythrocyte sedimentation rate (ESR). The aim of our study was to compare the ESR values by Monitor 100® against the standard Westergren method. This cross-sectional study was conducted at a Level I trauma care center on 200 patients. The samples taken were as per the recommendations charted out by International Council for Standardization in Hematology (ICSH) for comparing automated and manual Westergrens method. Bland and Altman statistical analysis was applied for evaluating Monitor 100® against the conventional Westergren method. The analysis revealed a low degree of agreement between the manual and automated methods especially for higher ESR values, mean difference -11.2 (95% limits of agreement, -46.3 to 23.9) and mean difference -13.4 (95% limits of agreement-58.9 to 32.1) for 1 and 2 hours, respectively. This discrepancy which is of clinical significance was less evident for ESR values in the normal range <25 mm/hour (-7.7 mean of difference; -18.9 to 3.5 limits of agreement). The fully automated system Monitor 100® for ESR measurement tends to underestimate the manual ESR readings. Hence it is recommended that a correction factor be applied for the range of ESR values while using this equipment. Further studies and validation experiments would be required.


Phenylsilver - an unexpected one-dimensional coordination polymer of silver(I) tetrads.
Herein we present the structures of two distinct one-dimensional coordination polymers of phenylsilver. Both polymers consist of phenylsilver tetrads inter-connected via short silver-silver contacts (either one or two). The underlying mechanism for the formation by transmetalation was validated by synthesising an analogous fluorinated derivative of phenylsilver.


Genetic variants of angiotensin II receptors and cardiovascular risk in hypertension.
Renin-angiotensin systems may mediate cardiovascular disease pathogenesis through a balance of actions of angiotensin II on (potentially proatherogenic) constitutive type 1 (AT1R) and (potentially antiatherogenic) inducible type 2 (AT2R) receptors. We explored such potential roles in a prospective candidate gene association study. Cardiovascular end points (fatal, nonfatal, and silent myocardial infarction and coronary artery bypass surgery/angioplasty) were documented among 2579 healthy UK men (mean age, 56.1+/-3.5 years; median follow-up, 10.1 years) genotyped for the AT1R1166A>C and the X chromosome located AT2R1675A>G and 3123C>A polymorphisms. Baseline characteristics, including blood pressure, were independent of genotype. The AT1R1166CC genotype was associated with relative cardiovascular risk (hazard ratio, 1.65 [1.05 to 2.59]; P=0.03) independent of blood pressure. Systolic blood pressure was associated with risk (P=0.0005), but this association was restricted to AT2R1675A allele carriers (P<0.00001), with G allele carriers protected from the risk associated with blood pressure (P=0.18). Hypertensive carriers with the AT2R1675A/3123A haplotype were at most risk, with 37.5% having an event. This is the first study to demonstrate an association of AT2R genotype with coronary risk, an effect that was confined to hypertensive subjects and supports the concept that the inducible AT2R is protective. Conversely, the AT1R1166CC genotype was associated with cardiovascular risk irrespective of blood pressure. These data are important to our understanding of the divergent role of angiotensin II acting at its receptor subtypes and coronary disease pathogenesis and for the development of future cardiovascular therapies.


Salmonella enterica serotype Enteritidis and eggs: a national epidemic in the United States.
Beginning in the 1970s, the incidence of Salmonella enterica serotype Enteritidis (SE) infection and the number of related outbreaks in the United States has increased dramatically. By 1994, SE was the most commonly reported Salmonella serotype, with an incidence of >10 laboratory-confirmed infections per 100,000 population in the Northeast. Intensive epidemiologic and laboratory investigations identified shell eggs as the major vehicle for SE infection in humans, and that the eggs had been internally contaminated by transovarian transmission of SE in the laying hen. Three key interventions aimed at preventing the contamination and growth of SE in eggs have included farm-based programs to prevent SE from being introduced into egg-laying flocks, early and sustained refrigeration of shell eggs, and education of consumers and food workers about the risk of consuming raw or undercooked eggs. Since 1996, the incidence of SE infection in humans has decreased greatly, although many cases and outbreaks due to SE contaminated eggs continue to occur.


SDC2 and TFPI2 Methylation in Stool Samples as an Integrated Biomarker for Early Detection of Colorectal Cancer.
Detection of aberrant methylated DNA in the stool is an effective early screening method for colorectal cancer (CRC). Previously, reporters identified that syndecan-2 (SDC2) and tissue factor pathway inhibitor 2 (TFPI2) were aberrantly methylated in most CRC tissues. However, the combined diagnostic role of them remains undefined. Our research aimed at probing the role and efficiency of the methylation status of SDC2 and TFPI2 in CRC early screening by using bioinformatics analysis and clinical stool sample validation. The promoter and CpG site methylation levels of SDC2 and TFPI2 and their correlation with clinicopathological characteristics of CRC were analyzed using UALCAN, Methsurv, and Wanderer. UCSC Xena was used to perform survival analyses. LinkedOmics was used to do functional network analysis. DNA was isolated and purified from stool, and quantitative methylation-specific PCR (qMSP) was applied to detect methylatedSDC2 and TFPI2. The results showed that promoter and most CpG site methylation levels of SDC2 and TFPI2 were significantly higher in CRC than in normal tissues. Moreover, SDC2 and TFPI2 methylation showed a positive correlation. Functional network analysis suggested that both methylated SDC2 and TFPI2 were involved in tumor cells' metabolic programs. Besides, there was a higher positive integrated detection rate in CRC (n=61) with a sensitivity of 93.4% and in adenoma (Ade) (n=16) with a sensitivity of 81.3% than normal with a specificity of 94.3% in stool samples. What is more, integration of methylated SDC2 and TFPI2 showed a higher sensitivity and Youden index than a single gene in detecting Adeor CRC. Our data indicate that SDC2 and TFPI2 were hypermethylated in CRC, and integrated detection of methylated SDC2 and TFPI2 in stool has the potential to be an effective and noninvasive tool of CRC early screening.


Access to the brain parenchyma using endovascular techniques and a micro-working channel.
OBJECTIVE Several older studies report a low risk for parenchymal access to the CNS by surgical techniques. In more recent studies, including those with post-puncture CT scans, there are indications that the risk of bleeding might approach 8%. New therapies, such as those that use viral vectors, modified mRNA, or cell transplantation, will probably warrant more parenchymal access to the CNS. Other minimally invasive routes might then be tempting to explore. This study was designed in 2 parts to address the possibility of using the endovascular route. The first aim was to test the ability to create a parenchymal micro-working channel to the CNS in macaque monkeys through the vessel wall. Second, the biocompatibility of a device-associated, detached, distal securing plug that was made of nitinol was investigated in swine for 1 year. METHODS Trans-vessel wall intervention in the middle cerebral artery and associated cerebral parenchyma was performed in 4 rhesus macaque monkeys using a full clinical angiography suite. A contrast agent and methylene blue were injected to test the working channel and then detached at the distal end to act as a securing plug through the vessel wall. One-year follow-ups were also performed using angiography and histological analysis in 10 swine with 24 implants that were distributed in the external carotid artery tree. RESULTS The cerebral interventions were performed without acute bleeding. Both the contrast agent and methylene blue were infused into the brain parenchyma and subarachnoidal space via the endovascular micro-working channel (7 injections in 4 animals). In the 1-year follow-up period, the implant that was left in the external carotid vessel wall in the swine was covered by the endothelium, which was followed by dislodgement just outside the blood vessel with thin capsule formation. No stenosis in the artery was detected on 1-year angiography. The animals showed normal behavior and blood sample results during the follow-up period. This is the first histological demonstration of nitinol biocompatibility when the implant is positioned through an arterial wall and indicates that the trans-vessel wall technique is not comparable with stent placement and its ability to induce intimal hyperplasia and restenosis. CONCLUSIONS This study demonstrates that the trans-vessel wall technique is applicable to brain intervention in macaque monkeys, providing a micro-working channel for delivery or sampling. The long-term follow-up study of the detached device in swine showed no clinical or biochemical complications and a normal angiography appearance.


Pulmonary capillary pressure and tissue perfusion: clinical implications during resuscitation from shock.
During shock resuscitation, a combination of fluids, vasopressors, vasodilators, and inotropes is administered in order to achieve a cardiac output or overall oxygen delivery as per guidelines of individual clinicians. The measurement of ventricular end-diastolic pressure allows a clinician to describe a therapeutic goal of optimum cardiac output response to changes in end-diastolic pressure. This concept has formed the backbone of resuscitative strategies in many forms of shock. Ventricular end-diastolic pressure is indirectly measured as the pulmonary artery occlusion pressure (PAOP) in critically ill patients with the use of a pulmonary artery catheter. Cytokines and other mediators may injure the pulmonary capillary endothelium which will affect the rate of leakage in the pulmonary capillaries. This may have important clinical implications in the therapy of shock in inflammatory states such as sepsis and the adult respiratory distress syndrome. Therefore, the true edema-forming pressure within the pulmonary bed is of considerable importance to the intensivist at the bedside. True pulmonary capillary pressure represents the midpoint of the capillary bed and is the hydrostatic pressure which directly drives the rate of pulmonary interstitial edema formation. During shock resuscitation in disorders in which vascular integrity may be impaired, the ability to measure pulmonary capillary pressure would be of great clinical benefit. It is impossible to directly measure pulmonary capillary hydrostatic pressure in the intact lung and, therefore, only indirect measurements are clinically possible. Numerous studies have demonstrated the lack of consistent relationship between the pulmonary capillary pressure, PAOP, pulmonary artery diastolic pressure, and the severity of acute lung injury. The assumption that PAOP, and thus left atrial pressure, is a good indirect measurement of pulmonary filtration pressure within the capillary bed is erroneous, in particular in the presence of increased resistance within the pulmonary venous bed between the capillaries and the left atrium, as may exist in disorders in which there is cytokine production. It is now clear that a significant gradient between pulmonary capillary pressure and PAOP may be present in inflammatory disorders which are not present in noninflammatory states, and that pulmonary capillary pressure may be measured at the bedside of critically ill patients. Bedside measurement of pulmonary capillary pressure may allow for added precision in our therapeutic goals in resuscitation from inflammatory shock. If further studies confirm the reliability and reproducibility of bedside measurement, pulmonary capillary pressure may become an invaluable part of the hemodynamic profile in the critically ill patient in shock.


Childhood abuse and vasomotor symptoms among midlife women.
Childhood maltreatment is related to adverse health outcomes. However, the relation of childhood maltreatment to the menopause transition-a universal transition for women often accompanied by troubling symptoms such as vasomotor symptoms-is relatively underexplored. This study tested whether childhood abuse and neglect are associated with menopausal vasomotor symptoms, utilizing both physiologic and prospective self-report measures of vasomotor symptoms. In all, 295 nonsmoking perimenopausal and postmenopausal women aged 40 to 60 years with and without vasomotor symptoms completed psychosocial measures, including the Child Trauma Questionnaire, ambulatory physiologic (sternal skin conductance) and self-report measurement of vasomotor symptoms during wake and sleep, and actigraphy measurement of sleep. Relationships between childhood abuse/neglect and vasomotor symptoms during wake and sleep were tested in linear regression models controlling for demographics, body mass index, and menopause stage. 44% of the sample reported abuse or neglect during childhood. Among women reporting vasomotor symptoms, childhood sexual or physical abuse was associated with more frequent physiologically-recorded vasomotor symptoms during sleep (sexual abuse: b [SE] = 1.45 [0.52], P = 0.006; physical abuse: b [SE] = 0.97 [0.47], P = 0.03) in multivariable models. Among these women, women with a physical or sexual abuse history had approximately 1.5 to 2-fold the number of sleep vasomotor symptoms than women without this history. Childhood abuse is associated with more frequent physiologically-detected vasomotor symptoms during sleep.


Not only the seed matters: Farmers' perceptions of sources for banana planting materials in Uganda.
The adoption of improved seed and other planting material in developing countries shows mixed results despite their potential to increase agricultural productivity. To arrive at a better understanding of the observed adoption rates, a lot of research is focused on finding the cultivars and variety traits that are attractive to farmers. Given smallholder farmers' seed sourcing practices are often influenced by social ties and cultural norms, it is also relevant to understand where and why farmers seek to acquire planting material. In this study, means-end chain analysis was applied to understand farmers' perceptions of formal and informal sources of banana planting material. Means-end chain analysis allows respondents to select and verbalize their own constructs to evaluate a product or service. These personally relevant constructs are subsequently linked to their personal goals via laddering interviews. We interviewed 31 Ugandan banana farmers from Western and Central region. Farmers associated formal sources mainly with improved cultivars, tissue culture plantlets and low levels of diversity. Informal seed sources were mostly associated with traditional cultivars, suckers and high levels of diversity. The goals farmers pursued while acquiring planting material, such as financial gains, food security, and to sustain and develop the household, were fairly similar among different groups of farmers. The means through which farmers aimed and preferred to pursue these goals differed and could be related to aspects such as gender, production scale and production goals. These differences among farmers preferences for particular sources indicate that not only cultivar traits should be tailored to farmers' preferences and needs, but also the characteristics of the sources from which farmers access planting material.


Pharmacokinetics of norfloxacin in chronic renal failure.
Norfloxacin is an antibacterial drug chiefly eliminated by the kidney and therefore useful in the treatment of urinary tract infections. To study its pharmacokinetics in chronic renal failure, we administered a single oral dose of 400 mg to 14 patients and 6 controls with normal renal function. Patients were divided into three groups according to the severity of renal failure. Norfloxacin was measured in serum and urine by bioassay. Serum half-life in controls was 3.87 hours and prolonged to 5.85 hours in group I (creatinine clearance 80-45 ml/min), 7.25 hours (p less than 0.05) in group II (creatinine clearance 44-20 ml/min) and 8.34 hours (p less than 0.01) in group III patients (creatinine clearance less than 20 ml/min). A good linear correlation between the elimination rate constant and creatinine clearance (r = 0.79, p less than 0.01) has been found. Total urinary excretion in the 72 hour period after administration achieves 40.4% in controls, falls to 23.5% (p less than 0.05), 15.6% (p less than 0.01) and 8.2% (p less than 0.01) of administered dose in groups I, II and III, respectively. Similarly, urinary concentrations decrease in all patient groups with respect to controls. Our data show that effective urinary antibacterial concentrations of norfloxacin after 400 mg single oral dose were obtained even in patients with severe renal failure. In these patients systemic accumulation after repeated dose administration is a probable event. Therefore, dosage adjustment is advisable in patients with creatinine clearance less than 20 ml/min, although it will inevitably cause lower urinary concentrations.


Amino-acid site variability among natural and designed proteins.
Computational protein design attempts to create protein sequences that fold stably into pre-specified structures. Here we compare alignments of designed proteins to alignments of natural proteins and assess how closely designed sequences recapitulate patterns of sequence variation found in natural protein sequences. We design proteins using RosettaDesign, and we evaluate both fixed-backbone designs and variable-backbone designs with different amounts of backbone flexibility. We find that proteins designed with a fixed backbone tend to underestimate the amount of site variability observed in natural proteins while proteins designed with an intermediate amount of backbone flexibility result in more realistic site variability. Further, the correlation between solvent exposure and site variability in designed proteins is lower than that in natural proteins. This finding suggests that site variability is too uniform across different solvent exposure states (i.e., buried residues are too variable or exposed residues too conserved). When comparing the amino acid frequencies in the designed proteins with those in natural proteins we find that in the designed proteins hydrophobic residues are underrepresented in the core. From these results we conclude that intermediate backbone flexibility during design results in more accurate protein design and that either scoring functions or backbone sampling methods require further improvement to accurately replicate structural constraints on site variability.


The risk of intracerebral haemorrhage with smoking. The Melbourne Risk Factor Study Group.
To determine whether smoking contributes to the risk of primary intracerebral haemorrhage (ICH) a case-control study was carried out on 331 consecutive cases of first-episode ICH as verified by computed tomography. Patients were age- and sex-matched to 331 community-based controls. Unlike other forms of stroke where smoking is an established risk factor, there was no increase in risk of ICH with smoking in this study (odds ratio, OR, 1.07, 95% confidence interval, CI, 0.631.81). Similar ratios were obtained for past and current smokers. Neither the amount smoked nor the duration of smoking were associated with ICH. Further investigation, however, revealed an interaction between smoking and hypertension on the risk of ICH that was similar regardless of the amount of cigarettes currently smoked and was largely seen to be a phenomenon in men (OR 8.13, 95% CI 2.0432.42). This interaction is a new finding, but the post-hoc nature of this analysis requires that it be further tested, preferably in a large prospective study.


[Birth weight and later nutritional status, cognitive development and job status: a critical revision].
The impact of prenatal nutritional status, assessed through birth weight (BW) and their effects in the short, medium and long-term on nutritional status, cognitive development and job status in the adult life, has been a problem of interest for several researchers; as regards, some of these report a positive and significant association between these variables and others do not find any relation. Children with insufficient, low or very low BW despite the early more deteriorate nutritional status should present higher risk for brain maturation, failure cognitive development and lowered head circumference which implies both lowered brain volume and intellectual development. In the short and medium-term, this situation damages the learning process at school-age, while in the long-term this might condition the quality of jobs. At present, the body of knowledge pinpoints that findings related to these associations is not conclusive verifying a great controversy in these matters. This review article has the purpose of analyzing the current evidence, in order to stimulate research about to these aspects which are relevant for child development and their future life.


Unveiling Lived Experiences: Exploring the Health and Lifestyle Effects of COVID-19 on Healthcare Workers.
The COVID-19 pandemic brought about significant life disruptions among healthcare workers (HCWs), including changes in weight, eating habits, and physical activity. This qualitative study sought to evaluate the initial and longitudinal effects of health habits among HCWs throughout the pandemic. Data were collected through Qualtrics surveys at three points over a 2-year period with questions asking participants (n = 234) to describe whether they experienced changes in weight, eating behaviors, and physical activity and why they believe these changes occurred. The open-ended responses were analyzed following the summative content analysis approach. Four key themes emerged: (1) problematic eating patterns and habits, (2) disruptions in physical activity, (3) alterations in work environment and schedule, and (4) declines in mental health. Respondent reflections highlight the immediate and long-term pandemic-related effects on weight status for some, attributed to alterations in routines and health habits. Other HCWs reported a "reset" or indicated their habits may have been initially disrupted but normalized or improved over the 2-year time span. Findings underscore the need for strategies that support the physical and mental health of healthcare workers.


Functional analysis of a novel GATA3 mutation in a family with the hypoparathyroidism, deafness, and renal dysplasia syndrome.
The hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome is an autosomal dominant disorder caused by mutations of a member of the GATA-binding family of transcription factors, GATA3. This dual zinc finger transcription factor binds DNA with its C-terminal zinc finger (ZnF2) and stabilizes this binding with its N-terminal zinc finger (ZnF1). ZnF1 also interacts with other zinc finger proteins, notably Friend of GATA (FOG). The HDR syndrome has been described in patients with mutations affecting both ZnF1 and ZnF2 domains; the former result in inefficient interaction with FOG, and the latter result in disruption of DNA binding. We report a patient with renal failure, hypoparathyroidism, and bilateral hearing loss. Assessment of family members indicated that the disease arose as a de novo mutation in her mother. Analysis of GATA3 in the family revealed a heterozygous missense mutation resulting in a nonconservative change of a single amino acid (R276P) in the ZnF1 domain. Functional analysis using dissociation electrophoretic mobility shift and yeast two-hybrid assays showed reduced binding affinity to the GATA motifs but normal interaction with FOG in vitro. These results are consistent with the predicted functions of human GATA3-ZnF1 from three-dimensional molecular modeling and with HDR being a result of GATA3 haploinsufficiency.


TRE-dependent transcription activation by JDP2-CHOP10 association.
The c-Jun dimerization protein 2, JDP2, is a member of the activating protein 1 (AP-1) family of transcription factors. Overexpression of JDP2 has been shown to result in repression of AP-1-dependent transcription and inhibition of cellular transformation. Other studies suggested that JDP2 may function as an oncogene. Here we describe the identification of CHOP10, a member of the CCAAT enhancer binding proteins, as a protein associating with JDP2. In contrast to the inhibition of transcription by JDP2, JDP2-CHOP complex strongly enhances transcription from promoters containing TPA response elements (TRE), but not from those containing cyclic AMP response elements (CRE). The association between JDP2 and CHOP10 involves the leucine zipper motifs of both proteins, whereas, the basic domain of CHOP10 contributes to the association of the JDP2-CHOP10 complex with the DNA. DNA binding of JDP2-CHOP complex is observed both in vitro and in vivo. Finally, overexpression of JDP2 results in increased cell viability following ER stress and counteracts CHOP10 pro-apoptotic activity. JDP2 expression may determine the threshold for cell sensitivity to ER stress. This is the first report describing TRE-dependent activation of transcription by JDP2 and thus may provide an explanation for the as yet unexplored oncogenic properties of JDP2.


Expression, not sequence, distinguishes miR-238 from its miR-239ab sister miRNAs in promoting longevity in Caenorhabditis elegans.
MicroRNAs (miRNAs) regulate gene expression by base-pairing to target sequences in messenger RNAs (mRNAs) and recruiting factors that induce translational repression and mRNA decay. In animals, nucleotides 2-8 at the 5' end of the miRNA, called the seed region, are often necessary and sometimes sufficient for functional target interactions. MiRNAs that contain identical seed sequences are grouped into families where individual members have the potential to share targets and act redundantly. A rare exception seemed to be the miR-238/239ab family in Caenorhabditis elegans, as previous work indicated that loss of miR-238 reduced lifespan while deletion of the miR-239ab locus resulted in enhanced longevity and thermal stress resistance. Here, we re-examined these potentially opposing roles using new strains that individually disrupt each miRNA sister. We confirmed that loss of miR-238 is associated with a shortened lifespan but could detect no longevity or stress phenotypes in animals lacking miR-239a or miR-239b, individually or in combination. Additionally, dozens of genes were mis-regulated in miR-238 mutants but almost no gene expression changes were detected in either miR-239a or miR-239b mutants compared to wild type animals. We present evidence that the lack of redundancy between miR-238 and miR-239ab is independent of their sequence differences; miR-239a or miR-239b could substitute for the longevity role of miR-238 when expressed from the miR-238 locus. Altogether, these studies disqualify miR-239ab as negative regulators of aging and demonstrate that expression, not sequence, dictates the specific role of miR-238 in promoting longevity.


Soy intake and risk of breast cancer in Asians and Asian Americans.
Evidence from case-control studies suggests, although not entirely consistently, that soy intake may protect against breast cancer. The designs and findings of studies conducted in Asian women living in Japan, Singapore, China, and the United States are reviewed. Because of the considerably higher intake of soy by native Asians than by Asian Americans living in California and Hawaii, these studies investigated different segments of the dose-response relation between soy intake and breast cancer risk. Data are not sufficient to determine the amount or frequency of soy intake effective in protecting against breast cancer. Of concern is that soy intake may be homogeneously high in Asia, making it difficult to identify differences in breast cancer risk between high and moderate daily consumers. In studies conducted in Asian Americans, it is difficult to be certain that soy intake is not a marker of other factors related to Western lifestyle that are causally associated with risk of breast cancer. Additional studies assessing the role of soy and breast cancer are needed. These studies should assess intake of all food sources of soy, considering portion size as well as other dietary and nondietary factors that may confound the soy-breast cancer association. A better understanding of the mechanisms whereby soy intake may influence the risk of breast cancer is also needed. Dietary intervention studies with soy will provide information on the acute effects of soy on endogenous hormone concentrations. Cross-sectional and longitudinal studies are necessary to investigate the longer-term relations between hormone concentrations and soy intake in women.


Beyond the Usual Suspects and Towards Politicisation: Immigration in Quebec's Party Manifestos, 1991-2018.
Immigration is increasingly politicised: issues surrounding immigration are more salient and party immigration positions are becoming more polarised. Literature on this topic has overwhelmingly focused on Europe and the USA, while identifying radical right parties and particular social dynamics as the main drivers of politicisation. In this paper, we consider a case that exhibits growing politicisation, yet does not fit existing measures and explanation of this phenomenon: Quebec, Canada. Considering party manifestos between 1991 and 2018, we ask: how have Quebec major political parties treated immigration policy issues over time? We argue that Quebec's immigration partisan space is organised by a liberal-restrictive immigration policy continuum and a jurisdictional axis. Mapping provincial political parties' positions, we document a stable clustering towards liberal policy positions but a clear cleavage on the jurisdictional axis. We show that the emergence in 2012 of a highly moderate centre right party-the Coalition Avenir Québec-broke with these established positions and drove a politicisation of the issue. These results demonstrate the importance of considering alternative drivers to explain the politicisation of immigration, as well as of paying attention to the politics of immigration in contexts where pro-immigration discourses dominate.


Effects of treadmill training with load addition on non-paretic lower limb on gait parameters after stroke: A randomized controlled clinical trial.
The addition of load on the non-paretic lower limb for the purpose of restraining this limb and stimulating the use of the paretic limb has been suggested to improve hemiparetic gait. However, the results are conflicting and only short-term effects have been observed. This study aims to investigate the effects of adding load on non-paretic lower limb during treadmill gait training as a multisession intervention on kinematic gait parameters after stroke. With this aim, 38 subacute stroke patients (mean time since stroke: 4.5 months) were randomly divided into two groups: treadmill training with load (equivalent to 5% of body weight) on the non-paretic ankle (experimental group) and treadmill training without load (control group). Both groups performed treadmill training during 30min per day, for two consecutive weeks (nine sessions). Spatiotemporal and angular gait parameters were assessed by a motion system analysis at baseline, post-training (at the end of 9days of interventions) and follow-up (40days after the end of interventions). Several post-training effects were demonstrated: patients walked faster and with longer paretic and non-paretic steps compared to baseline, and maintained these gains at follow-up. In addition, patients exhibited greater hip and knee joint excursion in both limbs at post-training, while maintaining most of these benefits at follow-up. All these improvements were observed in both groups. Although the proposal gait training program has provided better gait parameters for these subacute stroke patients, our data indicate that load addition used as a restraint may not provide additional benefits to gait training.


Computed tomography findings in COVID-19 and atypical pneumonia: a comparative study.
Computed tomography (CT) has an important role in the rapid diagnosis, treatment, and management of lower respiratory tract infections. This study aimed to explore different imaging characteristics between Coronavirus disease 2019 (COVID-19) and atypical pneumonia (non-COVID-19) on chest CT of patients admitted to the emergency department. CT features of 120 patients with positive Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by reverse transcriptase-polymerase chain reaction (RT-PCR) and 83 patients with negative SARS-CoV-2 by RT-PCR but positive respiratory tract sample test results for other respiratory pathogens were retrospectively evaluated, findings were recorded and compared between the two groups. Compared to non-COVID-19, COVID-19 patients were more likely to have a peripheral (60.5% vs. 23.8%, p < 0.001) and bilateral distribution (72.3% vs. 41.3%, p < 0.001), patchy consolidations (45% vs. 28.9%, p = 0.021), ground glass opacity (GGO) (94.2% vs. 83.1%, p = 0.011), crazy paving patterns (55% vs. 31.3%, p < 0.001); but less likely to have centrilobular nodules (15% vs. 62.7%, p < 0.001), pleural effusion (3.3% vs. 10.8%, p = 0.032), multifocal consolidations (7.5% vs. 21.7%, p = 0.003), and random distribution (1.7% vs. 46.3%, p < 0.001). There were significant differences between the CT patterns of patients with COVID-19 and other atypical pneumonia. The presence of patchy consolidations, GGO, crazy paving patterns with typical peripheral, bilateral distribution, and absence of centrilobular nodules, pleural effusion, and multifocal consolidations may help to differentiate COVID-19 from atypical pneumonia.


A systematic overview of adolescent suicide prevention programs.
The purpose of this systematic overview was to summarize evidence about the effectiveness of adolescent suicide prevention curricula programs. A comprehensive search of published and unpublished literature resulted in retrieval of 11 relevant studies. The findings, of this overview indicate that there is currently insufficient evidence to support curriculum-based suicide prevention programs. The evidence suggests that there may be both beneficial and harmful effects of the programs on students. In most studies, knowledge related to suicide improved as a result of the programs. However, studies found both beneficial and harmful effects on attitudes related to suicide. One study found in increase in hopelessness and maladaptive coping for males following the intervention. The literature suggests that more broadly based comprehensive school health programs should be evaluated for their effectiveness in addressing the determinants of adolescent risk behaviour.


Activation of the alternative pathway of complement in psoriatic lesional skin.
The complement system may play an important role in the inflammatory reaction of psoriasis. While the classical pathway of the complement has been shown to be activated in psoriasis, there have been few reports on the involvement of the alternative pathway in the inflammatory reactions in psoriasis. Complement fragments, C4d and Bb, are released at the time of the classical and alternative pathway activation, respectively. The presence of the C4d or Bb fragment, therefore, denotes a preceding complement activation through the classical and/or alternative pathway. In the present study, we have measured the levels of C4d and Bb in psoriatic scale extracts using enzyme immunoassay techniques. The scales of these dermatoses contained higher levels of C4d and Bb detectable by enzyme immunoassay than those in the stratum corneum of noninflammatory skin. These results suggest that the alternative as well as the classical pathway of complement are activated in psoriatic lesional skin.


Tuning the self-assembly and luminescence properties of lanthanide coordination polymers by ligand design.
Two new structure-related tripodal ligands featuring salicylamide pendant arms, 1,3,5-tris{[(2'-furfurylaminoformyl)phenoxyl]methyl}-2,4,6-trimethylbenzene (L(I)) and 1,1,1-tris{[(2'-furfurylaminoformyl)phenoxyl]methyl}ethane (L(II)) have been designed and synthesized with the ultimate aim of self-assembling lanthanide polymers with interesting luminescent properties. Among two series of Ln(III) nitrate complexes (Ln = Pr, Nd, Sm, Eu, Gd, Tb or Dy) which have been characterized by elemental analyses, XRD, TGA and IR spectra, three new coordination polymers have been determined by X-ray diffraction analysis. The coordination polymer type {[Ln(NO(3))(3)(L(I))].nH(2)O}(n) possesses an unusual ladderlike double chain which can be further connected through pi-pi stacking interactions constructing a three-dimensional supramolecular structure. In contrast, the coordination polymer type {[Ln(NO(3))(3)(L(II))].nCH(3)OH}(n) displays a (3,3)-connected puckered two-dimensional net with 4.8(2) topological notation. The photophysical properties of the Sm, Eu, Tb and Dy complexes at room temperature are investigated. The present work substantiates the claim that the supramolecular structure as well as the luminescent properties of the coordination polymer can be tuned by varying either the backbone group or the terminal group of the organic ligand.


Cabozantinib for progressive metastatic medullary thyroid cancer: a review.
Medullary thyroid cancer is uncommon and patients typically present with advanced disease. Treatment options for patients with progressive, metastatic medullary thyroid cancer had been limited until recently. Tyrosine kinase inhibitors have garnered increasing interest in this subset of patients. The US Food and Drug Administration recently approved cabozantinib, a tyrosine kinase inhibitor, after promising results were shown in a large Phase III clinical trial. This review summarizes the clinical pharmacology, clinical trials, and safety data for cabozantinib and concludes with a discussion of possible future directions for the treatment of medullary thyroid cancer.


Precise laser poration to control drug delivery into and through human nail.
Drug treatment of diseases of the human nail remains a difficult challenge; topical therapy, in particular, is limited by very poor transport of active agents across the nail itself. The objective of this research was to examine the potential of controlled, and fibre-optic delivered, femtosecond laser light pulses to provide new pathways and opportunities for drug access to targets within and beneath the nail plate. Optical, confocal fluorescence and scanning electron microscopies demonstrated partial and complete laser poration of human nail samples, with the energy per pore and the exposure duration being the key modulating parameters that determined the extent of ablation achieved. Parallel measurements of the penetration of a model drug across laser-treated nails showed that complete poration resulted in essentially complete circumvention of the diffusion barrier, an array of 100 pores in 0.2cm2 area of nail permitting a 103-fold increase in initial drug uptake. Partial ablation of the nail created pores that extended to a range of depths; the nail material adjacent to the ablated area was rendered porous in appearance presumably due to local thermal perturbation of the nail structure. These openings offer, as a result, potential sites in which topical drug formulations might be sequestered post-poration and from which slow, sustained delivery of the active agent into and through the nail may be envisaged.


Automatic implantable defibrillators.
The automatic implantable defibrillator was developed in the USA by Mirowski. This device automatically detects ventricular fibrillation or ventricular tachycardia and delivers a low energy (30 J) shock directly to the myocardium via patch electrodes on the epicardial surface or through a combination of a pericardial patch and an endocardial wire. These devices are being implanted in large numbers in the USA and represent a major contribution to the management and refractory ventricular arrhythmias. Unfortunately their high cost has limited their use in this country despite impressive clinical results.


Indirect chelatometric determination of piperazine in some pharmaceutical preparations.
An indirect complexometric method is described for the determination of piperazine alone or in its pharmaceutical preparations. The method involves treatment of the aqueous solution of piperazine with carbon disulphide, ammonia solution and a known excess of cupric acetate, filtration of the formed precipitate and determination of the excess cupric acetate with EDTA using PAR as indicator.


Non-reductive continental naturalism in the contemporary humanities: Working with Hélène Metzger's philosophical reflections.
This article engages with the philosophical reflections of the French historian of science Hélène Metzger (1886-1944) in order to develop a vocabulary for understanding the rise of non-reductive Continental naturalism in the contemporary humanities. The bibliography of current naturalist approaches in the arts and the human sciences is still in the making, but it is altogether clear that the trend is not scientist or historicist or relativist. This epistemological diagnosis refers us to Metzger, who found herself surrounded with the logical positivism of the Wiener Kreis, on the one hand, and the historicism of her French colleagues, on the other, as well as with the infiltration of the history of science by a chronological empiricism. In this article I will take the most recent book of Vicki Kirby - Quantum Anthropologies: Life at Large from 2011 - as an exemplary case of non-reductive Continental naturalist scholarship in the humanities today and by reading it through the concepts of Metzger, I will demonstrate how this type of research leads to refreshing insights in what constitutes positive humanities knowledge and what is the role of the a priori in the field.


Career tracks in academic family medicine: issues and approaches.
There are a number of possible career tracks available to individuals interested in full-time teaching in either community-based family practice residency programs or medical school departments of family medicine. These include various kinds of experience in residency an fellowship training and clinical practice. Full-time faculty may be involved in any of four major areas: patient care, teaching, research, and administration. The balance of an individual's responsibilities in each of these areas varies considerably based upon the teaching setting and the individual's strengths, interests, and career stage. Continued personal growth and career development of faculty of the joint responsibility of individual faculty members and directors of chairman of residency programs and departments. Diversity is required among family practice faculty in order for the multiple needs of academic family medicine to be effectively addressed in patient care, teaching, and research. The first generation of family practice faculty have been principally clinicians, teachers, and administrators. There is now a particular need for faculty who can integrate clinical, teaching, and research skills.


Very low birth weight infants: effects of brain growth during infancy on intelligence quotient at 3 years of age.
To evaluate the role of postnatal growth on IQ at 3 years of age, 139 appropriate for gestational age, very low birth weight infants (less than 1.5 kg) born in 1977 and 1978 were studied at 40 weeks (term), and at 8, 20, and 33 months (corrected) of age. Weight, height, and head circumference were measured at each age, neurologic status was measured at 20 months, and Stanford Binet IQ at 33 months. Multiple regression analysis revealed that head circumference at 8 months of age is the best growth predictor of IQ at 3 years of age. Path analysis was performed to measure the effects of biologic and social factors measured earlier in life on IQ at 3 years. These factors explained 43% of the variance in IQ at 3 years of age. Head circumference at 8 months had a direct effect on IQ at 3 years, controlling for all other variables in the model. Neonatal risk had an indirect effect via head circumference. Neurologic impairment had direct and indirect (via head circumference) effects; race and socioeconomic status had direct effects on IQ but no effects on growth at 8 months of age. Thus, brain growth at 8 months significantly influenced 3 year IQ at 3 years of age among very low birth weight infants, even when medical and sociodemographic variables were controlled.


The employment interview: avoiding the traps with effective strategies.
Selecting well-qualified employees has become increasingly difficult for employers. Legal complaints citing discrimination and negligent hiring have made interviewers and organizations more vulnerable than ever. Also, predicted declines in the skill levels of employees will continue to make it difficult for interviewers to select applicants who best match job requirements. These difficulties can be traced to a variety of traps that often lead to poor hiring decisions. The authors describe the traps interviewers face and highlight actions that can be taken by interviewers to avoid these traps and to improve the effectiveness of their interviewing and employee selection decisions.


Entrustment Evidence Used by Expert Gynecologic Surgical Teachers to Determine Residents' Autonomy.
To identify entrustment evidence used by expert gynecologic surgical teachers to determine obstetrics and gynecology residents' level of autonomy in the operating room. A qualitative interview study was undertaken from March to November 2016. Four selection criteria were used to define and purposefully sample expert gynecologic surgical teachers across the United States to represent all four geographic regions. All interviews were audio-recorded and transcribed. We applied the Framework Method of content analysis. Transcripts were iteratively analyzed and emergent themes identified. Twenty-seven expert gynecologic surgical teachers from 15 institutions across the United States participated in 30-minute interviews. We identified four domains of entrustment evidence (resident characteristics, medical knowledge, technical performance, and "beyond current surgical case") commonly reported by expert gynecologic surgical teachers to determine residents' autonomy as well as the particular evidence associated with expert gynecologic surgical teachers' determination of resident autonomy at two decision-making points (surgical time-out and taking over certain intraoperative steps) in the operating room. Onsite direct observation and conversation were two common methods used by expert gynecologic surgical teachers to obtain this evidence. Entrustment evidence from resident characteristics, medical knowledge, and technical performance domains and from "beyond current surgical case" was commonly used by expert gynecologic surgical teachers to determine residents' autonomy. Our findings provide a potential framework for designing educational interventions that aim to increase residents' readiness for autonomy and entrustment in the operating room.


Persistence of a Wolbachia infection frequency cline in Drosophila melanogaster and the possible role of reproductive dormancy.
Field populations of arthropods are often polymorphic for Wolbachia but the factors maintaining intermediate Wolbachia frequencies are generally not understood. In Drosophila melanogaster, Wolbachia frequencies are highly variable across the globe. We document the persistence of a Wolbachia infection frequency cline in D. melanogaster populations from eastern Australia across at least 20 years, with frequencies generally high in the tropics but lower in cool temperate regions. The results are interpreted using a model of frequency dynamics incorporating cytoplasmic incompatibility (CI), imperfect maternal transmission and Wolbachia effects on fitness. Clinal variation is less pronounced in eastern North America which may reflect annual recolonization at higher latitudes. Limited samples from Africa from latitudes matching our tropical and subtropical samples from Australia and North America show comparably high infection frequencies, but some equatorial samples show lower frequencies. Adult dormancy across cold periods may contribute to the Australian Wolbachia cline. Infected flies exposed to cold conditions for an extended period had reduced fecundity and viability, an effect not evident in unexposed controls. These fitness costs may contribute to the relatively low Wolbachia frequencies in Australian temperate areas; whereas different processes, including CI induced by young males, may contribute to higher frequencies in tropical locations.


Relationship between phylogenetic groups, genotypic clusters, and virulence gene profiles of Escherichia coli strains from diverse human and animal sources.
Escherichia coli strains in water may originate from various sources, including humans, farm and wild animals, waterfowl, and pets. However, potential human health hazards associated with E. coli strains present in various animal hosts are not well known. In this study, E. coli strains from diverse human and animal sources in Minnesota and western Wisconsin were analyzed for the presence of genes coding for virulence factors by using multiplex PCR and biochemical reactions. Of the 1,531 isolates examined, 31 (2%) were found to be Shiga toxin-producing E. coli (STEC) strains. The majority of these strains, which were initially isolated from the ruminants sheep, goats, and deer, carried the stx(1c) and/or stx(2d), ehxA, and saa genes and belonged to E. coli phylogenetic group B1, indicating that they most likely do not cause severe human diseases. All the STEC strains, however, lacked eae. In contrast, 26 (1.7%) of the E. coli isolates examined were found to be potential enteropathogenic E. coli (EPEC) strains and consisted of several intimin subtypes that were distributed among various human and animal hosts. The EPEC strains belonged to all four phylogenetic groups examined, suggesting that EPEC strains were relatively widespread in terms of host animals and genetic background. Atypical EPEC strains, which carried an EPEC adherence factor plasmid, were identified among E. coli strains from humans and deer. DNA fingerprint analyses, done using the horizontal, fluorophore-enhanced repetitive-element, palindromic PCR technique, indicated that the STEC, potential EPEC, and non-STEC ehxA-positive E. coli strains were genotypically distinct and clustered independently. However, some of the potential EPEC isolates were genotypically indistinguishable from nonpathogenic E. coli strains. Our results revealed that potential human health hazards associated with pathogenic E. coli strains varied among the animal hosts that we examined and that some animal species may harbor a greater number of potential pathogenic strains than other animal species.


Cardiopulmonary resuscitation and hypothermia.
Current basic life support (BLS) protocols do not address the physiologic effects of accidental hypothermia in prehospital care. The extreme levels of bradycardia, bradypnea, and peripheral vasoconstriction that often accompany profound hypothermia may complicate the accurate diagnosis of cardiopulmonary arrest in the unmonitored patient. Although CPR is indicated in the truly pulseless, apneic victim of hypothermia, chest compressions may convert nonpalpable but adequately perfusing sinus bradycardia to ventricular fibrillation. This dilemma had led to disagreement among clinicians and researchers in hypothermia about prehospital care protocols for the severely hypothermic patient. This article reviews the controversy and recommends the application of a normal BLS protocol to hypothermic patients presenting in apparent cardiopulmonary arrest.


The human histone gene cluster at the D6S105 locus.
The sequences and organization of the histone genes in the histone gene cluster at the chromosomal marker D6S105 have been determined by analyzing the Centre d'Etude du Polymorphisme Humain yeast artificial chromosome (YAC) 964f1. The insert of the YAC was subcloned in cosmids. In the established contig of the histone-gene-containing cosmids, 16 histone genes and 2 pseudogenes were identified: one H1 gene (H1.5), five H2A genes, four H2B genes and one pseudogene of H2B, three H3 genes, and three H4 genes plus one H4 pseudogene. The cluster extends about 80 kb with a nonordered arrangement of the histone genes. The dinucleotide repeat polymorphic marker D6S105 was localized at the telomeric end of this histone gene cluster. Almost all human histone genes isolated until now have been localized within this histone gene cluster and within the previously described region of histone genes, about 2 Mb telomeric of the newly described cluster or in a small group of histone genes on chromosome 1. We therefore conclude that the data presented here complete the set of human histone genes. This now allows the general organization of the human histone gene complement to be outlined on the basis of a compilation of all known histone gene clusters and solitary histone genes.


The relationship between school-based health centers, rates of early dismissal from school, and loss of seat time.
This study sought to understand the relationship between school-based health centers (SBHCs) and academic outcomes such as early dismissal and loss of seat time (the time students are available in school to learn or to access support services). A quasi-experimental research design was used to compare rates of early dismissal and loss of seat time between students who received SBHC and traditional school nursing services and students who received only traditional school nursing services. This study was a secondary data analysis of 764 "walk-in" visits during a 3-week period in 2 urban high schools in western New York state. Both schools provided school nursing services, and 1 of the 2 offered the option to enroll in an SBHC. SBHCs significantly reduced the number of early dismissals from school (p = .013) in a comparison with students who received school nursing services alone. Students not enrolled in an SBHC lost 3 times as much seat time as students enrolled in an SBHC. Race, gender, age, poverty status, and presence of a preexisting illness did not influence these findings. These findings suggest that SBHCs have a direct impact on educational outcomes such as attendance. Recommendations for further research include replication of this study to increase confidence in its findings and using early dismissal and loss of seat time as indicators of attendance to measure other health outcomes related to SBHCs and school nursing.


[Concepts in face lifts. State of the art].
A brief review of the historical development of face-lifting is followed by a description of the current concepts in order to judge the general aspects of modern face-lifting better; composite face-lifting, sub-SMAS, extended face-lifting, video-assisted face-lifting, and laser-assisted skin rejuvenation are discussed in detail. The results show that there is no such thing as a general face-lift. Each of the methods described has a specific indication, but no longer under the general outdated aspect that the face should be "tightened". Rather, modern rhytidectomy should be understood as anatomical repositioning of ptotic soft tissue, especially the nasolabial fat pad. Individual problem zones must be included in the subtitle surgical strategy, which involves individualized techniques and weighing the benefits and risks.


APC/C--the master controller of origin licensing?
DNA replication must be tightly controlled to prevent initiation of a second round of replication until mitosis is complete. So far, components of the pre-replicative complex (Cdt1, Cdc6 and geminin) were considered key players in this regulation. In a new study, Machida and Dutta have shown that depletion of Emi1 caused cells to replicate their DNA more than once per cell cycle 1. This effect was dependent on the ability of Emi1 to inhibit the APC/C. In addition to its role in regulating entry into mitosis, oscillation of APC/C activity regulates pre-RC formation: high APC/C activity in late M/G1 allows pre-RC formation and low APC/C activity in S/G2 prevents pre-RC formation for a second time thereby preventing rereplication. Each redundant pathway to prevent rereplication is dependent on regulating one of the pre-RC components, and all of the pathways are co-regulated by Emi1 through the APC/C. In this commentary we discuss how this new role of Emi1 adds to our understanding of the regulation of replication initiation. We also review the literature to analyze whether APC/C has a role in regulating endoreduplication (a normal state of polyploidy in some differentiated cells). Similarly a role of premature APC/C activation in genomic instability of tumors is discussed.


The use of mHealth to deliver tailored messages reduces reported energy and fat intake.
Evidence supports the role of feedback in reinforcing motivation for behavior change. Feedback that provides reinforcement has the potential to increase dietary self-monitoring and enhance attainment of recommended dietary intake. The aim of this study was to examine the impact of daily feedback (DFB) messages, delivered remotely, on changes in dietary intake. This was a secondary analysis of the Self- Monitoring And Recording using Technology (SMART) Trial, a single-center, 24-month randomized clinical trial of behavioral treatment for weight loss. Participants included 210 obese adults (mean body mass index, 34.0 kg/m²) who were randomized to either a paper diary (PD), personal digital assistant (PDA), or PDA plus daily tailored feedback messages (PDA + FB). To determine the role of daily tailored feedback in dietary intake, we compared the self-monitoring with DFB group (DFB group; n = 70) with the self-monitoring without DFB group (no-DFB group, n = 140). All participants received a standard behavioral intervention for weight loss. Self-reported changes in dietary intake were compared between the DFB and no-DFB groups and were measured at baseline and at 6, 12, 18, and 24 months. Linear mixed modeling was used to examine percentage changes in dietary intake from baseline. Compared with the no-DFB group, the DFB group achieved a larger reduction in energy (-22.8% vs -14.0%; P = .02) and saturated fat (-11.3% vs -0.5%; P = .03) intake and a trend toward a greater decrease in total fat intake (-10.4% vs -4.7%; P = .09). There were significant improvements over time in carbohydrate intake and total fat intake for both groups (P values < .05). Daily tailored feedback messages designed to target energy and fat intake and delivered remotely in real time using mobile devices may play an important role in the reduction of energy and fat intake.


Tetrameric assembly of hGBP1 is crucial for both stimulated GMP formation and antiviral activity.
Interferon-γ inducible human guanylate binding protein-1 (hGBP1) shows a unique characteristic that hydrolyses GTP to a mixture of GDP and GMP through successive cleavages, with GMP being the major product. Like other large GTPases, hGBP1 undergoes oligomerization upon substrate hydrolysis, which is essential for the stimulation of activity. It also exhibits antiviral activity against many viruses including hepatitis C. However, which oligomeric form is responsible for the stimulated activity leading to enhanced GMP formation and its influence on antiviral activity, are not properly understood. Using mutant and truncated proteins, our data indicate that transition-state-induced tetramerization is associated with higher rate of GMP formation. This is supported by chimaeras that are defective in both tetramerization and enhanced GMP formation. Unlike wild-type protein, chimaeras did not show allosteric interactions, indicating that tetramerization and enhanced GMP formation are allosterically coupled. Hence, we propose that after the cleavage of the first phosphoanhydride bond GDP·Pi-bound protein dimers transiently associate to form a tetramer that acts as an allosteric switch for higher rate of GMP formation. Biochemical and biophysical studies reveal that sequential conformational changes and interdomain communications regulate tetramer formation via dimer. Our studies also show that overexpression of the mutants, defective in tetramer formation in Rep2a cells do not inhibit proliferation of hepatitis C virus, indicating critical role of a tetramer in the antiviral activity. Thus, the present study not only highlights the importance of hGBP1 tetramer in stimulated GMP formation, but also demonstrates its role in the antiviral activity against hepatitis C virus.


Designing Electrostatic Interactions via Polyelectrolyte Monomer Sequence.
Charged polymers are ubiquitous in biological systems because electrostatic interactions can drive complicated structure formation and respond to environmental parameters such as ionic strength and pH. In these systems, function emerges from sophisticated molecular design; for example, intrinsically disordered proteins leverage specific sequences of monomeric charges to control the formation and function of intracellular compartments known as membraneless organelles. The role of a charged monomer sequence in dictating the strength of electrostatic interactions remains poorly understood despite extensive evidence that sequence is a powerful tool biology uses to tune soft materials. In this article, we use a combination of theory, experiment, and simulation to establish the physical principles governing sequence-driven control of electrostatic interactions. We predict how arbitrary sequences of charge give rise to drastic changes in electrostatic interactions and correspondingly phase behavior. We generalize a transfer matrix formalism that describes a phase separation phenomenon known as "complex coacervation" and provide a theoretical framework to predict the phase behavior of charge sequences. This work thus provides insights into both how charge sequence is used in biology and how it could be used to engineer properties of synthetic polymer systems.


Heterozygous germline mutations in the CBL tumor-suppressor gene cause a Noonan syndrome-like phenotype.
RAS signaling plays a key role in controlling appropriate cell responses to extracellular stimuli and participates in early and late developmental processes. Although enhanced flow through this pathway has been established as a major contributor to oncogenesis, recent discoveries have revealed that aberrant RAS activation causes a group of clinically related developmental disorders characterized by facial dysmorphism, a wide spectrum of cardiac disease, reduced growth, variable cognitive deficits, ectodermal and musculoskeletal anomalies, and increased risk for certain malignancies. Here, we report that heterozygous germline mutations in CBL, a tumor-suppressor gene that is mutated in myeloid malignancies and encodes a multivalent adaptor protein with E3 ubiquitin ligase activity, can underlie a phenotype with clinical features fitting or partially overlapping Noonan syndrome (NS), the most common condition of this disease family. Independent CBL mutations were identified in two sporadic cases and two families from among 365 unrelated subjects who had NS or suggestive features and were negative for mutations in previously identified disease genes. Phenotypic heterogeneity and variable expressivity were documented. Mutations were missense changes altering evolutionarily conserved residues located in the RING finger domain or the linker connecting this domain to the N-terminal tyrosine kinase binding domain, a known mutational hot spot in myeloid malignancies. Mutations were shown to affect CBL-mediated receptor ubiquitylation and dysregulate signal flow through RAS. These findings document that germline mutations in CBL alter development to cause a clinically variable condition that resembles NS and that possibly predisposes to malignancies.


Hyperexcitability and epilepsy associated with disruption of the mouse neuronal-specific K-Cl cotransporter gene.
Four genes encode electroneutral, Na+-independent, K-Cl cotransporters. KCC2, is exclusively expressed in neurons where it is thought to drive intracellular Cl- to low concentrations and shift the reversal potential for Cl- conductances such as GABA(A) or glycine receptor channels, thus participating in the postnatal development of inhibitory mechanisms in the brain. Indeed, expression of the cotransporter is low at birth and increases postnatally, at a time when the intracellular Cl- concentration in neurons decreases and gamma-aminobutyric acid switches its effect from excitatory to inhibitory. To assert the significance of KCC2 in neuronal function, we disrupted the mouse gene encoding this neuronal-specific K-Cl cotransporter. We demonstrate that animals deficient in KCC2 exhibit frequent generalized seizures and die shortly after birth. We also show upregulation of Fos, the product of the immediate early gene c-fos, and the significant loss of parvalbumin-positive interneurons, both indicative of brain injury. The regions most affected are the hippocampus and temporal and entorhinal cortices. Extracellular field potential measurements in the CA1 hippocampus exhibited hyperexcitability. Application of picrotoxin, a blocker of the GABA(A) receptor, further increased hyperexcitability in homozygous hippocampal sections. Pharmacological treatment of pups showed that diazepam relieved the seizures while phenytoin prevented them between postnatal ages P4-P12. Finally, we demonstrate that adult heterozygote animals show increased susceptibility for epileptic seizure and increased resistance to the anticonvulsant effect of propofol. Taken together, these results indicate that KCC2 plays an important role in controlling CNS excitability during both postnatal development and adult life.


Gender Differences in All-Cause Mortality after Acute Myocardial Infarction: Evidence for a Gender-Age Interaction.
Gender difference studies in mortality after acute myocardial infarction (AMI) have shown inconsistent results. A total of 13,104 patients from the KAMIR-NIH between November 2011 and December 2015 were classified into young (n = 3837 [29.3%]) and elderly (n = 9267 [70.7%]) patients. For the study, women <65 and men <55 years of age were considered "young". In the adjusted model of the entire cohort, there was no significant difference in three-year all-cause mortality between women and men (17.8% vs. 10.3%; adjusted hazard ratio [HR], 0.953; 95% confidence interval [CI], 0.799-1.137). However, when the entire cohort was subdivided into two age groups, young women showed an 84.3% higher mortality rate than young men (adjusted HR, 1.843; 95% CI, 1.098-3.095). Contrariwise, elderly women patients had a 20.4% lower hazard of mortality compared with elderly men (adjusted HR, 0.796; 95% CI, 0.682-0.929). The interaction of gender with age was significant, even after multiple adjustments (adjusted p for interaction = 0.003). The purpose of this study was to assess whether gender differences depend on the patients' age. Based on our analysis, higher mortality of young women remains even in the contemporary era of AMI. A better understanding of the mechanisms underlying these differences is warranted.


The distribution and 'in vivo' phase variation status of haemoglobin receptors in invasive meningococcal serogroup B disease: genotypic and phenotypic analysis.
Two haemoglobin-binding proteins, HmbR and HpuAB, contribute to iron acquisition by Neisseria meningitidis. These receptors are subject to high frequency, reversible switches in gene expression--phase variation (PV)--due to mutations in homopolymeric (poly-G) repeats present in the open reading frame. The distribution and PV state of these receptors was assessed for a representative collection of isolates from invasive meningococcal disease patients of England, Wales and Northern Ireland. Most of the major clonal complexes had only the HmbR receptor whilst the recently expanding ST-275-centred cluster of the ST-269 clonal complex had both receptors. At least one of the receptors was in an 'ON' configuration in 76.3% of the isolates, a finding that was largely consistent with phenotypic analyses. As PV status may change during isolation and culture of meningococci, a PCR-based protocol was utilised to confirm the expression status of the receptors within contemporaneously acquired clinical specimens (blood/cerebrospinal fluid) from the respective patients. The expression state was confirmed for all isolate/specimen pairs with <15 tract repeats indicating that the PV status of these receptors is stable during isolation. This study therefore establishes a protocol for determining in vivo PV status to aid in determining the contributions of phase variable genes to invasive meningococcal disease. Furthermore, the results of the study support a putative but non-essential role of the meningococcal haemoglobin receptors as virulence factors whilst further highlighting their vaccine candidacy.